Synchrotron Radiolysis and Mass Spectrometry: A New Approach to Research on the Actin Cytoskeleton

Jing Qu Guan, Steven C. Almo, Mark R. Chance

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Hydroxyl radicals generated from millisecond exposure of aqueous solutions to synchrotron X-rays react with proteins to yield stable oxidative modifications of solvent-accessible amino acid side chains. Following proteolysis, HPLC/MS analysis is performed to quantitate the side chain reactivities, and MS/MS analysis is used to identify the modification site(s). Side chain reactivity is shown to be correlated with solvent accessibility; thus the method provides detailed site-specific information about protein structure. The application of these techniques to the study of the actin cytoskeleton is described in detail, including defining the binding sites of monomeric actin with gelsolin segment-1, the actin monomer binding surface on cofilin, the divalent cation-dependent structure changes of monomeric actin, and the conformational changes associated with actin filamentous assembly.

Original languageEnglish (US)
Pages (from-to)221-229
Number of pages9
JournalAccounts of Chemical Research
Volume37
Issue number4
DOIs
StatePublished - Apr 2004

Fingerprint

Radiolysis
Synchrotrons
Mass spectrometry
Actins
Actin Depolymerizing Factors
Gelsolin
Proteolysis
Divalent Cations
Hydroxyl Radical
Proteins
Monomers
Binding Sites
Amino Acids
X rays

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Synchrotron Radiolysis and Mass Spectrometry : A New Approach to Research on the Actin Cytoskeleton. / Guan, Jing Qu; Almo, Steven C.; Chance, Mark R.

In: Accounts of Chemical Research, Vol. 37, No. 4, 04.2004, p. 221-229.

Research output: Contribution to journalArticle

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