RGD Reference Report - The local electrostatic environment determines cysteine reactivity of tubulin. - Rat Genome Database

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The local electrostatic environment determines cysteine reactivity of tubulin.

Authors: Britto, PJ  Knipling, L  Wolff, J 
Citation: Britto PJ, etal., J Biol Chem 2002 Aug 9;277(32):29018-27.
RGD ID: 730142
Pubmed: PMID:12023292   (View Abstract at PubMed)
DOI: DOI:10.1074/jbc.M204263200   (Journal Full-text)

Of the 20 cysteines of rat brain tubulin, some react rapidly with sulfhydryl reagents, and some react slowly. The fast reacting cysteines cannot be distinguished with [14C]iodoacetamide, N-[(14)C]ethylmaleimide, or IAEDANS ([5-((((2-iodoacetyl)amino)ethyl)amino) naphthalene-1-sulfonic acid]), since modification to mole ratios 1 cysteine/dimer always leads to labeling of 6-7 cysteine residues. These have been identified as Cys-305alpha, Cys-315alpha, Cys-316alpha, Cys-347alpha, Cys-376alpha, Cys-241beta, and Cys-356beta by mass spectroscopy and sequencing. This lack of specificity can be ascribed to reagents that are too reactive; only with the relatively inactive chloroacetamide could we identify Cys-347alpha as the most reactive cysteine of tubulin. Using the 3.5-A electron diffraction structure, it could be shown that the reactive cysteines were within 6.5 A of positively charged arginines and lysines or the positive edges of aromatic rings, presumably promoting dissociation of the thiol to the thiolate anion. By the same reasoning the inactivity of a number of less reactive cysteines could be ascribed to inhibition of modification by negatively charged local environments, even with some surface-exposed cysteines. We conclude that the local electrostatic environment of cysteine is an important, although not necessarily the only, determinant of its reactivity.

Objects referenced in this article
Gene Tuba1a tubulin, alpha 1A Rattus norvegicus

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