Myosin Subfragment 1 and Structural Elements of G-Actin

Effects of S-1(A2) on Sequences 39-52 and 61-69 in Subdomain 2 of G-Actin

Theresa Chen, Missak Haigentz, Emil Reisler

Research output: Contribution to journalArticle

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Abstract

The effect of myosin on the structure of two sequences on G-actin, a loop between residues 39 and 52 and a segment between residues 61 and 69 from the NH2-terminus, was probed by limited proteolytic digestions of G-actin in the presence of the myosin subfragment 1 isozyme S-1(A2). Under the experimental conditions of this work, no polymerization of actin was induced by S-1(A2) [Chen & Reisler (1991) Biochemistry 30, 4546-4552]. S-1(A2) did not change the rates of subtilisin and chymotryptic digestion of G-actin at loop 39-52. In contrast to this, the second protease-sensitive region on G-actin, segment 61-69, was protected strongly by S-1(A2) from tryptic cleavage. The minor if any involvement of loop 39-52 in S-l binding was confirmed by determining the binding constants of S-1(A2) for pyrene-labeled G-actin (1.2 X 106 M-1), subtilisin-cleaved pyrenyl G-actin (0.3 X 106 M-1), and DNase I-pyrenyl G-actin complexes (0.3 X 106 M-1). Consistent with this, the activity of DNase I, which binds to actin loop 39-52 [Kabsch et al. (1990) Nature 347, 37-44], was inhibited almost equally well by actin in the presence and absence of S-1(A2). These results confirm the observation that DNase I and S-1(A2) bind to distinct sites on actin [Bettache et al. (1990) Biochemistry 29, 9085-9091] and demonstrate myosin-induced changes in segment 61—69 of G-actin.

Original languageEnglish (US)
Pages (from-to)2941-2946
Number of pages6
JournalBiochemistry
Volume31
Issue number11
DOIs
StatePublished - Mar 1 1992
Externally publishedYes

Fingerprint

Myosin Subfragments
varespladib methyl
Actins
Deoxyribonuclease I
Subtilisin
Biochemistry
Myosins
Digestion
Polymerization
Isoenzymes
Peptide Hydrolases

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

@article{3b3e294d884b47bf94a1647ab1b1e310,
title = "Myosin Subfragment 1 and Structural Elements of G-Actin: Effects of S-1(A2) on Sequences 39-52 and 61-69 in Subdomain 2 of G-Actin",
abstract = "The effect of myosin on the structure of two sequences on G-actin, a loop between residues 39 and 52 and a segment between residues 61 and 69 from the NH2-terminus, was probed by limited proteolytic digestions of G-actin in the presence of the myosin subfragment 1 isozyme S-1(A2). Under the experimental conditions of this work, no polymerization of actin was induced by S-1(A2) [Chen & Reisler (1991) Biochemistry 30, 4546-4552]. S-1(A2) did not change the rates of subtilisin and chymotryptic digestion of G-actin at loop 39-52. In contrast to this, the second protease-sensitive region on G-actin, segment 61-69, was protected strongly by S-1(A2) from tryptic cleavage. The minor if any involvement of loop 39-52 in S-l binding was confirmed by determining the binding constants of S-1(A2) for pyrene-labeled G-actin (1.2 X 106 M-1), subtilisin-cleaved pyrenyl G-actin (0.3 X 106 M-1), and DNase I-pyrenyl G-actin complexes (0.3 X 106 M-1). Consistent with this, the activity of DNase I, which binds to actin loop 39-52 [Kabsch et al. (1990) Nature 347, 37-44], was inhibited almost equally well by actin in the presence and absence of S-1(A2). These results confirm the observation that DNase I and S-1(A2) bind to distinct sites on actin [Bettache et al. (1990) Biochemistry 29, 9085-9091] and demonstrate myosin-induced changes in segment 61—69 of G-actin.",
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Myosin Subfragment 1 and Structural Elements of G-Actin : Effects of S-1(A2) on Sequences 39-52 and 61-69 in Subdomain 2 of G-Actin. / Chen, Theresa; Haigentz, Missak; Reisler, Emil.

In: Biochemistry, Vol. 31, No. 11, 01.03.1992, p. 2941-2946.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Myosin Subfragment 1 and Structural Elements of G-Actin

T2 - Effects of S-1(A2) on Sequences 39-52 and 61-69 in Subdomain 2 of G-Actin

AU - Chen, Theresa

AU - Haigentz, Missak

AU - Reisler, Emil

PY - 1992/3/1

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N2 - The effect of myosin on the structure of two sequences on G-actin, a loop between residues 39 and 52 and a segment between residues 61 and 69 from the NH2-terminus, was probed by limited proteolytic digestions of G-actin in the presence of the myosin subfragment 1 isozyme S-1(A2). Under the experimental conditions of this work, no polymerization of actin was induced by S-1(A2) [Chen & Reisler (1991) Biochemistry 30, 4546-4552]. S-1(A2) did not change the rates of subtilisin and chymotryptic digestion of G-actin at loop 39-52. In contrast to this, the second protease-sensitive region on G-actin, segment 61-69, was protected strongly by S-1(A2) from tryptic cleavage. The minor if any involvement of loop 39-52 in S-l binding was confirmed by determining the binding constants of S-1(A2) for pyrene-labeled G-actin (1.2 X 106 M-1), subtilisin-cleaved pyrenyl G-actin (0.3 X 106 M-1), and DNase I-pyrenyl G-actin complexes (0.3 X 106 M-1). Consistent with this, the activity of DNase I, which binds to actin loop 39-52 [Kabsch et al. (1990) Nature 347, 37-44], was inhibited almost equally well by actin in the presence and absence of S-1(A2). These results confirm the observation that DNase I and S-1(A2) bind to distinct sites on actin [Bettache et al. (1990) Biochemistry 29, 9085-9091] and demonstrate myosin-induced changes in segment 61—69 of G-actin.

AB - The effect of myosin on the structure of two sequences on G-actin, a loop between residues 39 and 52 and a segment between residues 61 and 69 from the NH2-terminus, was probed by limited proteolytic digestions of G-actin in the presence of the myosin subfragment 1 isozyme S-1(A2). Under the experimental conditions of this work, no polymerization of actin was induced by S-1(A2) [Chen & Reisler (1991) Biochemistry 30, 4546-4552]. S-1(A2) did not change the rates of subtilisin and chymotryptic digestion of G-actin at loop 39-52. In contrast to this, the second protease-sensitive region on G-actin, segment 61-69, was protected strongly by S-1(A2) from tryptic cleavage. The minor if any involvement of loop 39-52 in S-l binding was confirmed by determining the binding constants of S-1(A2) for pyrene-labeled G-actin (1.2 X 106 M-1), subtilisin-cleaved pyrenyl G-actin (0.3 X 106 M-1), and DNase I-pyrenyl G-actin complexes (0.3 X 106 M-1). Consistent with this, the activity of DNase I, which binds to actin loop 39-52 [Kabsch et al. (1990) Nature 347, 37-44], was inhibited almost equally well by actin in the presence and absence of S-1(A2). These results confirm the observation that DNase I and S-1(A2) bind to distinct sites on actin [Bettache et al. (1990) Biochemistry 29, 9085-9091] and demonstrate myosin-induced changes in segment 61—69 of G-actin.

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