Effects of binding factors on structural elements in F-actin

Damon Scoville; John D Stamm; Christian Altenbach; Alexander Shvetsov; Kaveh Kokabi; Peter A Rubenstein; Wayne L Hubbell; Emil Reisler

doi:10.1021/bi801649j

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Effects of binding factors on structural elements in F-actin

Journal article

Open access

Peer reviewed

Effects of binding factors on structural elements in F-actin

Damon Scoville, John D Stamm, Christian Altenbach, Alexander Shvetsov, Kaveh Kokabi, Peter A Rubenstein, Wayne L Hubbell and Emil Reisler

Biochemistry (Easton), Vol.48(2), pp.370-378

01/20/2009

DOI: 10.1021/bi801649j

PMCID: PMC3133778

PMID: 19113841

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https://www.ncbi.nlm.nih.gov/pmc/articles/3133778View

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Abstract

Understanding the dynamics of the actin filament is essential to a detailed description of their interactions and role in the cell. Previous studies have linked the dynamic properties of actin filaments (F-actin) to three structural elements contributing to a hydrophobic pocket, namely, the hydrophobic loop, the DNase I binding loop, and the C-terminus. Here, we examine how these structural elements are influenced by factors that stabilize or destabilize F-actin, using site-directed spin-labeled (SDSL) electron paramagnetic resonance (EPR), fluorescence, and cross-linking techniques. Specifically, we employ cofilin, an actin destabilizing protein that binds and severs filaments, and phalloidin, a fungal toxin that binds and stabilizes F-actin. We find that cofilin shifts both the DNase I binding loop and the hydrophobic loop away from the C-terminus in F-actin, as demonstrated by weakened spin-spin interactions, and alters the environment of spin probes on residues of these two loops. In contrast, although phalloidin strongly stabilizes F-actin, it causes little or no local change in the environment of the loop residues. This indicates that the stabilizing effect of phalloidin is achieved mainly through constraining structural fluctuations in F-actin and suggests that factors and interactions that control these fluctuations have an important role in the cytoskeleton dynamics.

Mutation

Amino Acid Sequence

Temperature

Actins - isolation & purification

Cross-Linking Reagents - chemistry

Electron Spin Resonance Spectroscopy

Protein Structure, Secondary

Saccharomyces cerevisiae - genetics

Actins - ultrastructure

Fluorescent Dyes - metabolism

Models, Molecular

Molecular Sequence Data

Saccharomyces cerevisiae - chemistry

Actins - genetics

2-Naphthylamine - analogs & derivatives

Actin Depolymerizing Factors - metabolism

Saccharomyces cerevisiae - cytology

Phalloidine - metabolism

Disulfides - chemistry

2-Naphthylamine - metabolism

Actins - chemistry

Protein Conformation

Catalysis

Details

Title: Subtitle: Effects of binding factors on structural elements in F-actin
Creators: Damon Scoville - Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California, Los Angeles, California 90095, USA
John D Stamm
Christian Altenbach
Alexander Shvetsov
Kaveh Kokabi
Peter A Rubenstein
Wayne L Hubbell
Emil Reisler
Resource Type: Journal article
Publication Details: Biochemistry (Easton), Vol.48(2), pp.370-378
DOI: 10.1021/bi801649j
PMID: 19113841
PMCID: PMC3133778
NLM abbreviation: Biochemistry
ISSN: 0006-2960
eISSN: 1520-4995
Publisher: United States
Grant note: R01 GM077190 / NIGMS NIH HHS GM 077190 / NIGMS NIH HHS R01 GM033689 / NIGMS NIH HHS T32 EY07026 / NEI NIH HHS T32 EY007026 / NEI NIH HHS EY 05216 / NEI NIH HHS R01 GM077190-31 / NIGMS NIH HHS GM 33689 / NIGMS NIH HHS R01 GM077190-30 / NIGMS NIH HHS R37 EY005216 / NEI NIH HHS R01 EY005216 / NEI NIH HHS
Language: English
Date published: 01/20/2009
Academic Unit: Stead Family Department of Pediatrics; Biochemistry and Molecular Biology; Internal Medicine
Record Identifier: 9984024520902771

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