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A Nucleotide State-sensing Region on Actin
Journal article   Open access   Peer reviewed

A Nucleotide State-sensing Region on Actin

Dmitri S Kudryashov, Elena E Grintsevich, Peter A Rubenstein and Emil Reisler
The Journal of biological chemistry, Vol.285(33), pp.25591-25601
08/13/2010
DOI: 10.1074/jbc.M110.123869
PMCID: PMC2919123
PMID: 20530485
url
https://doi.org/10.1074/jbc.M110.123869View
Published (Version of record) Open Access

Abstract

The nucleotide state of actin (ATP, ADP-P i , or ADP) is known to impact its interactions with other actin molecules upon polymerization as well as with multiple actin binding proteins both in the monomeric and filamentous states of actin. Recently, molecular dynamics simulations predicted that a sequence located at the interface of subdomains 1 and 3 (W-loop; residues 165–172) changes from an unstructured loop to a β-turn conformation upon ATP hydrolysis (Zheng, X., Diraviyam, K., and Sept, D. (2007) Biophys. J. 93, 1277–1283). This region participates directly in the binding to other subunits in F-actin as well as to cofilin, profilin, and WH2 domain proteins and, therefore, could contribute to the nucleotide sensitivity of these interactions. The present study demonstrates a reciprocal communication between the W-loop region and the nucleotide binding cleft on actin. Point mutagenesis of residues 167, 169, and 170 and their site-specific labeling significantly affect the nucleotide release from the cleft region, whereas the ATP/ADP switch alters the fluorescence of probes located in the W-loop. In the ADP-P i state, the W-loop adopts a conformation similar to that in the ATP state but different from the ADP state. Binding of latrunculin A to the nucleotide cleft favors the ATP-like conformation of the W-loop, whereas ADP-ribosylation of Arg-177 forces the W-loop into a conformation distinct from those in the ADP and ATP-states. Overall, our experimental data suggest that the W-loop of actin is a nucleotide sensor, which may contribute to the nucleotide state-dependent changes in F-actin and nucleotide state-modulated interactions of both G- and F-actin with actin-binding proteins.
 Protein Structure and Folding Actin Kabiramide C Nucleotide Cytoskeleton Protein Conformation Protein Structure WH2 Domain ADP ATP SpvB

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