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PMID: 3911945 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The use of actin labelled with N-(1-pyrenyl)iodoacetamide to study the interaction of actin with myosin subfragments and troponin/tropomyosin.

The Biochemical journal ·Vol. 232 ·No. 2 ·1985-12-01 ·Pages 343-9

Criddle AH, Geeves MA, Jeffries T

Abstract

A pyrene label attached to Cys-374 of actin has been shown to be a useful probe for monitoring the interaction of actin with myosin subfragments [Kouyama & Mihashi (1981) Eur. J. Biochem. 114, 33-38]. We report that the presence of this label decreases the affinity of actin for myosin subfragment 1 by less than a factor of 2. The rate of actin binding is unaffected by the label and the dissociation rate is increased by up to a factor of 2. Both the rate of actin binding to, and the rate of actin dissociation from, heavy meromyosin show two phases when monitored by pyrene fluorescence. Thin filiments reconstituted from pyrene-labelled actin show a 5% increase in pyrene fluorescence on binding Ca2+.

MeSH Terms
Actins/metabolism Adenosine Triphosphate/pharmacology Animals Binding Sites Fluorescent Dyes Iodoacetamide/analogs & derivatives Iodoacetates Kinetics Muscle Proteins/metabolism Myosin Subfragments/metabolism Myosins/metabolism Peptide Fragments/metabolism Rabbits Tropomyosin/metabolism Troponin/metabolism
Chemicals
Actins Fluorescent Dyes Iodoacetates Muscle Proteins Myosin Subfragments Peptide Fragments Tropomyosin Troponin N-(1-pyrenyl)iodoacetamide Adenosine Triphosphate Myosins Iodoacetamide
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Criddle A H
Geeves M A
Jeffries T
References (28)
28 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2320343
Published
1985-12-01
Pages
343-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1152885
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2320343 PMC Full Text View journal details
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