Cofilin induced conformational changes in F-actin expose subdomain 2 to proteolysis

Andras Muhlrad, Dmitry Kudryashov, Y. Michael Peyser, Andrey A. Bobkov, Steven C. Almo, Emil Reisler

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Cofilin/ADF affects strongly the structure of actin filaments and especially the intermolecular contacts of the DNase I binding loop (D-loop) in subdomain 2. In G-actin, the D-loop is cleaved by subtilisin between Met47 and Gly48, while in F-actin this cleavage is inhibited. Here, we report that yeast cofilin, which is resistant to both subtilisin and trypsin, accelerates greatly the rate of subtilisin cleavage of this loop in F-actin at pH 6.8 and at pH 8.0. Similarly, cofilin accelerates strongly the tryptic cleavage in F-actin of loop 60-69 in subdomain 2, at Arg62 and Lys68. The acceleration of the loops' proteolysis cannot be attributed to an increased treadmilling of F-actin for the following reasons: (i) the rate of subtilisin cleavage is independent of pH between pH 6.8 and 8.0, unlike F-actin depolymerization, which is pH-dependent; (ii) at high concentrations of protease the cleavage rate of F-actin in the presence of cofilin is faster than the rate of monomer dissociation from the pointed end of TRC-labeled F-actin, which limits the rate of treadmilling; and (iii) cofilin also accelerates the rate of subtilisin cleavage of F-actin in which the treadmilling is blocked by interprotomer cross-linking of the D-loop to the C terminus on an adjacent protomer. This suggests a substantial flexibility of the D-loop in the cross-linked F-actin. The increased cleavage rates of the D-loop and loop 60-69 reveal extensive exposure of subdomain 2 in F-actin to proteolytic enzymes by cofilin.

Original languageEnglish (US)
Pages (from-to)1559-1567
Number of pages9
JournalJournal of Molecular Biology
Volume342
Issue number5
DOIs
StatePublished - Oct 1 2004

Fingerprint

Actin Depolymerizing Factors
Proteolysis
Actins
Subtilisin
Peptide Hydrolases
Deoxyribonuclease I
Protein Subunits
Actin Cytoskeleton
Trypsin

Keywords

  • actin
  • ADF
  • cofilin
  • DNase I binding loop
  • protein-protein interaction

ASJC Scopus subject areas

  • Virology

Cite this

Cofilin induced conformational changes in F-actin expose subdomain 2 to proteolysis. / Muhlrad, Andras; Kudryashov, Dmitry; Michael Peyser, Y.; Bobkov, Andrey A.; Almo, Steven C.; Reisler, Emil.

In: Journal of Molecular Biology, Vol. 342, No. 5, 01.10.2004, p. 1559-1567.

Research output: Contribution to journalArticle

Muhlrad, A, Kudryashov, D, Michael Peyser, Y, Bobkov, AA, Almo, SC & Reisler, E 2004, 'Cofilin induced conformational changes in F-actin expose subdomain 2 to proteolysis', Journal of Molecular Biology, vol. 342, no. 5, pp. 1559-1567. https://doi.org/10.1016/j.jmb.2004.08.010
Muhlrad, Andras ; Kudryashov, Dmitry ; Michael Peyser, Y. ; Bobkov, Andrey A. ; Almo, Steven C. ; Reisler, Emil. / Cofilin induced conformational changes in F-actin expose subdomain 2 to proteolysis. In: Journal of Molecular Biology. 2004 ; Vol. 342, No. 5. pp. 1559-1567.
@article{5045d681fce3427984aec83fd88176e8,
title = "Cofilin induced conformational changes in F-actin expose subdomain 2 to proteolysis",
abstract = "Cofilin/ADF affects strongly the structure of actin filaments and especially the intermolecular contacts of the DNase I binding loop (D-loop) in subdomain 2. In G-actin, the D-loop is cleaved by subtilisin between Met47 and Gly48, while in F-actin this cleavage is inhibited. Here, we report that yeast cofilin, which is resistant to both subtilisin and trypsin, accelerates greatly the rate of subtilisin cleavage of this loop in F-actin at pH 6.8 and at pH 8.0. Similarly, cofilin accelerates strongly the tryptic cleavage in F-actin of loop 60-69 in subdomain 2, at Arg62 and Lys68. The acceleration of the loops' proteolysis cannot be attributed to an increased treadmilling of F-actin for the following reasons: (i) the rate of subtilisin cleavage is independent of pH between pH 6.8 and 8.0, unlike F-actin depolymerization, which is pH-dependent; (ii) at high concentrations of protease the cleavage rate of F-actin in the presence of cofilin is faster than the rate of monomer dissociation from the pointed end of TRC-labeled F-actin, which limits the rate of treadmilling; and (iii) cofilin also accelerates the rate of subtilisin cleavage of F-actin in which the treadmilling is blocked by interprotomer cross-linking of the D-loop to the C terminus on an adjacent protomer. This suggests a substantial flexibility of the D-loop in the cross-linked F-actin. The increased cleavage rates of the D-loop and loop 60-69 reveal extensive exposure of subdomain 2 in F-actin to proteolytic enzymes by cofilin.",
keywords = "actin, ADF, cofilin, DNase I binding loop, protein-protein interaction",
author = "Andras Muhlrad and Dmitry Kudryashov and {Michael Peyser}, Y. and Bobkov, {Andrey A.} and Almo, {Steven C.} and Emil Reisler",
year = "2004",
month = "10",
day = "1",
doi = "10.1016/j.jmb.2004.08.010",
language = "English (US)",
volume = "342",
pages = "1559--1567",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "5",

}

TY - JOUR

T1 - Cofilin induced conformational changes in F-actin expose subdomain 2 to proteolysis

AU - Muhlrad, Andras

AU - Kudryashov, Dmitry

AU - Michael Peyser, Y.

AU - Bobkov, Andrey A.

AU - Almo, Steven C.

AU - Reisler, Emil

PY - 2004/10/1

Y1 - 2004/10/1

N2 - Cofilin/ADF affects strongly the structure of actin filaments and especially the intermolecular contacts of the DNase I binding loop (D-loop) in subdomain 2. In G-actin, the D-loop is cleaved by subtilisin between Met47 and Gly48, while in F-actin this cleavage is inhibited. Here, we report that yeast cofilin, which is resistant to both subtilisin and trypsin, accelerates greatly the rate of subtilisin cleavage of this loop in F-actin at pH 6.8 and at pH 8.0. Similarly, cofilin accelerates strongly the tryptic cleavage in F-actin of loop 60-69 in subdomain 2, at Arg62 and Lys68. The acceleration of the loops' proteolysis cannot be attributed to an increased treadmilling of F-actin for the following reasons: (i) the rate of subtilisin cleavage is independent of pH between pH 6.8 and 8.0, unlike F-actin depolymerization, which is pH-dependent; (ii) at high concentrations of protease the cleavage rate of F-actin in the presence of cofilin is faster than the rate of monomer dissociation from the pointed end of TRC-labeled F-actin, which limits the rate of treadmilling; and (iii) cofilin also accelerates the rate of subtilisin cleavage of F-actin in which the treadmilling is blocked by interprotomer cross-linking of the D-loop to the C terminus on an adjacent protomer. This suggests a substantial flexibility of the D-loop in the cross-linked F-actin. The increased cleavage rates of the D-loop and loop 60-69 reveal extensive exposure of subdomain 2 in F-actin to proteolytic enzymes by cofilin.

AB - Cofilin/ADF affects strongly the structure of actin filaments and especially the intermolecular contacts of the DNase I binding loop (D-loop) in subdomain 2. In G-actin, the D-loop is cleaved by subtilisin between Met47 and Gly48, while in F-actin this cleavage is inhibited. Here, we report that yeast cofilin, which is resistant to both subtilisin and trypsin, accelerates greatly the rate of subtilisin cleavage of this loop in F-actin at pH 6.8 and at pH 8.0. Similarly, cofilin accelerates strongly the tryptic cleavage in F-actin of loop 60-69 in subdomain 2, at Arg62 and Lys68. The acceleration of the loops' proteolysis cannot be attributed to an increased treadmilling of F-actin for the following reasons: (i) the rate of subtilisin cleavage is independent of pH between pH 6.8 and 8.0, unlike F-actin depolymerization, which is pH-dependent; (ii) at high concentrations of protease the cleavage rate of F-actin in the presence of cofilin is faster than the rate of monomer dissociation from the pointed end of TRC-labeled F-actin, which limits the rate of treadmilling; and (iii) cofilin also accelerates the rate of subtilisin cleavage of F-actin in which the treadmilling is blocked by interprotomer cross-linking of the D-loop to the C terminus on an adjacent protomer. This suggests a substantial flexibility of the D-loop in the cross-linked F-actin. The increased cleavage rates of the D-loop and loop 60-69 reveal extensive exposure of subdomain 2 in F-actin to proteolytic enzymes by cofilin.

KW - actin

KW - ADF

KW - cofilin

KW - DNase I binding loop

KW - protein-protein interaction

UR - http://www.scopus.com/inward/record.url?scp=4444357229&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4444357229&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2004.08.010

DO - 10.1016/j.jmb.2004.08.010

M3 - Article

VL - 342

SP - 1559

EP - 1567

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 5

ER -