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An isobaric quantitative crosslinking mass spectrometry technology for studying conformational and structural changes in proteins and protein complexes

ABSTRACT: Experimental determination of protein conformational states is crucial to understand the mechanism of protein function and how function is altered in disease. We introduce a novel, simple and compact, isobaric crosslinker design for studying conformational and structural changes using quantitative mass spectrometry.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Jie Luo

LAB HEAD: Jeff Ranish

PROVIDER: PXD035939 | Pride | 2024-11-13

REPOSITORIES: Pride

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			Action	DRS
	ExpII.inter1.txt	Txt
	ExpII.intra1.txt	Txt
	Exp_I.inter1.txt	Txt
	Exp_I.intra1.txt	Txt
	JL102921_110821_RPB4_C1.mzXML	Mzxml

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Isobaric crosslinking mass spectrometry technology for studying conformational and structural changes in proteins and complexes

Project description:Dynamic conformational and structural changes in proteins and protein complexes play a central and ubiquitous role in the regulation of protein function, yet it is very challenging to study these changes, especially for large protein complexes, under physiological conditions. Here we introduce a novel isobaric crosslinker, Qlinker, for studying conformational and structural changes in proteins and protein complexes using quantitative crosslinking mass spectrometry (qCLMS). Qlinkers are small and simple, amine-reactive molecules with an optimal extended distance of ~10 Å which use MS2 reporter ions for relative quantification of Qlinker-modified peptides derived from different samples. We synthesized the 2-plex Q2linker and showed that the Q2linker can provide quantitative crosslinking data that pinpoints key conformational and structural changes in biosensors, binary and ternary complexes composed of the general transcription factors TBP, TFIIA, and TFIIB, and RNA polymerase II (pol II) complexes.

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