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PMID: 6863518 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Pyrene actin: documentation of the validity of a sensitive assay for actin polymerization.

Journal of muscle research and cell motility ·Vol. 4 ·No. 2 ·1983-04-00 ·Pages 253-62

Cooper JA, Walker SB, Pollard TD

Abstract

The fluorescence of pyrene-labelled actin is much higher after polymerization. We have characterized in detail the polymerization properties of pyrene actin and report that native and pyrene actin are identical using the following criteria: (1) the time course of polymerization; (2) the elongation rate constants; (3) the intrinsic viscosity; and (4) the critical concentration. Native and pyrene actin copolymerize. Fluorescence of polymerized pyrene actin is 7-10 times higher than monomer. The fluorescent signal is proportional to polymer weight concentration and is insensitive to filament length distribution. Bleaching can be minimized by appropriate filters to allow continuous monitoring of signal. Measurements do not influence polymerization kinetics. This establishes that pyrene actin fluorescence is a valid assay for actin polymerization that is more sensitive than any other current assay.

MeSH Terms
Actins/metabolism Biopolymers Calcium/pharmacology Kinetics Magnesium/pharmacology Pyrenes Spectrometry, Fluorescence Viscosity
Chemicals
Actins Biopolymers Pyrenes pyrene Magnesium Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Cooper J A
Walker S B
Pollard T D
References (17)
17 references, click to expand
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Article Info
Journal
Journal of muscle research and cell motility
Abbr.
J Muscle Res Cell Motil
ISSN
0142-4319
DOI
10.1007/BF00712034
Published
1983-04-00
Pages
253-62
Language
English
Region
Netherlands
NLM ID
8006298
Subset
IM
Grants
NIGMS NIH HHS · GM 7039 · United States
NIGMS NIH HHS · GM-26338 · United States
External Links
PubMed source DOI: 10.1007/BF00712034 View journal details
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