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PMID: 7272443 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Fluorescence photobleaching recovery in solutions of labeled actin.

Biophysical journal ·Vol. 35 ·No. 2 ·1981-08-00 ·Pages 351-64

Lanni F, Taylor DL, Ware BR

Abstract

We have demonstrated that the technique of fluorescence photobleaching recovery (FPR) can be used to examine the state of a single component in complex self-assembling macromolecular systems. Polymerization of actin, initiated by addition of salt or Mg+2 to a low-ionic-strength solution of G-actin, has been observed by sequential measurement of FPR with the aid of fluorescein-labeled actin. Solutions of actin which had been labeled using 5-iodoacetamido fluorescein (5-IAF) showed anomalous recovery of fluorescence above the initial value, which indicates a photoinduced increase in local polymerization. No such anomaly was observed with actin that had been labeled with fluorescein isothiocyanate (FITC). The FPR data are directly interpretable in terms of the fraction of labeled protein that is immobilized in the supramolecular assembly and in terms of the average diffusion coefficient of the mobile fraction. Our data are consistent with the "treadmill" model of actin polymerization, in that they show that actin is present under polymerizing conditions either as a high polymer or as monomer or low oligomer. We believe that the FPR technique can be applied to the study of many types of reconstituted motile or cytoskeletal systems in vitro or in vivo.

MeSH Terms
Actins/metabolism Animals Macromolecular Substances Mathematics Muscles/metabolism Photolysis Rabbits Solutions Spectrometry, Fluorescence
Chemicals
Actins Macromolecular Substances Solutions
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lanni F
Taylor D L
Ware B R
References (51)
51 references, click to expand
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(81)84794-7
Published
1981-08-00
Pages
351-64
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1327527
Subset
IM
External Links
PubMed source DOI: 10.1016/S0006-3495(81)84794-7 PMC Full Text View journal details
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