Source 1primary paper2020Journal of the American Society for Mass Spectrometry
Toolkit/ultraviolet photodissociation
ultraviolet photodissociation
Also known as: 213 nm ultraviolet photodissociation, UVPD
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
213 nm ultraviolet photodissociation (UVPD) is a top-down mass spectrometry fragmentation method used to characterize covalent insulin dimers. In the cited study, it fragmented cross-links and nearby backbone bonds and, together with multistage analysis or complementary fragmentation, supported identification of cross-link composition and residue-level cross-link sites.
Usefulness & Problems
Why this is useful
This method is useful for structural characterization of covalently cross-linked protein dimers by providing informative fragmentation of both the cross-link and adjacent peptide backbone. In insulin dimers, it enabled cross-link site assignment in at least one dimer class at the MS2 level and contributed to chemical composition analysis when combined with EThcD.
Source:
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Source:
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Source:
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Problem solved
It addresses the analytical problem of localizing and chemically characterizing covalent cross-links in intact protein dimers by top-down mass spectrometry. The cited work specifically used it to distinguish and map cross-links in Fe2+-induced and UV light-induced insulin dimers.
Problem links
Need precise spatiotemporal control with light input
Derived213 nm ultraviolet photodissociation (UVPD) is a top-down mass spectrometry fragmentation method used to characterize covalent insulin dimers. In the cited study, it enabled fragmentation of cross-links and nearby backbone bonds and contributed to identifying cross-link composition and, with multistage analysis, residue-level cross-link sites.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
backbone bond cleavagebackbone bond cleavageHeterodimerizationphotofragmentation of cross-linksphotofragmentation of cross-linksultraviolet photodissociationultraviolet photodissociationTechniques
Functional AssayTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: multi component delivery burdenimplementation constraint: spectral hardware requirementoperating role: sensorswitch architecture: multi componentswitch architecture: recruitment
The method uses 213 nm ultraviolet irradiation within a top-down mass spectrometry workflow. The cited implementation included MS2 analysis and, in some cases, multistage or combined fragmentation with EThcD to improve cross-link composition and site assignment.
MS2 UVPD was not sufficient for cross-link site identification in the UV light-induced insulin dimer, indicating context-dependent performance. Evidence is limited to a single 2020 study on insulin dimers, so broader generalizability across proteins and cross-link chemistries is not established here.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Residue-level identification of cross-link sites between chains was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds.
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
Top-down mass spectrometry workflows can identify cross-link types and sites in covalent insulin dimers induced by Fe2+ incubation or UV light stress.
we here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
At the MS2 level, EThcD efficiently cleaved interchain disulfide bonds in insulin dimers to reveal cross-link connectivities between chains.
On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the Fe2+-induced insulin dimer, Phe1 from both B-chains were cross-linked through a -CH2- linkage.
In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-.
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
In the UV light-induced insulin dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain.
in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain
Approval Evidence
1 source4 linked approval claimsfirst-pass slug ultraviolet-photodissociation
213 nm ultraviolet photodissociation (UVPD)
Source:
comparative method performancesupports
UVPD at MS2 provided cross-link site identification for the Fe2+-induced insulin dimer without MS3, but MS2 was not sufficient for cross-link site identification in the UV light-induced dimer.
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
Source:
mechanistic explanationsupports
The UV chromophoric side chain of Phe1 was implicated in the Fe2+-induced dimer cross-link, explaining why UVPD-MS2 effectively fragmented the cross-link and nearby backbone bonds.
The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds.
Source:
method capabilitysupports
Combined EThcD and 213 nm UVPD facilitated identification of the chemical composition of cross-links in insulin dimers.
The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links.
Source:
method conclusionsupports
HCD, EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.
Source:
Comparisons
Source-backed strengths
In the Fe2+-induced insulin dimer, UVPD at MS2 was sufficient for cross-link site identification without requiring MS3. The study also reported that combined EThcD and 213 nm UVPD facilitated identification of cross-link chemical composition, indicating complementarity with other fragmentation modes.
Source:
UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer.
Compared with electron-electron double resonance spectroscopy
ultraviolet photodissociation and electron-electron double resonance spectroscopy address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: heterodimerization; same primary input modality: light
Compared with electron-transfer/higher-energy collision dissociation
ultraviolet photodissociation and electron-transfer/higher-energy collision dissociation address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: heterodimerization, ultraviolet photodissociation; same primary input modality: light
Strengths here: looks easier to implement in practice.
Compared with top-down mass spectrometry
ultraviolet photodissociation and top-down mass spectrometry address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: heterodimerization; same primary input modality: light
Ranked Citations
- 1.