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PMID: 8460118 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Charge-reversion mutagenesis of Dictyostelium actin to map the surface recognized by myosin during ATP-driven sliding motion.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 6 ·1993-03-15 ·Pages 2127-31

Johara M, Toyoshima YY, Ishijima A, Kojima H, Yanagida T, Sutoh K

Abstract

Amino acid residues D24/D25, E99/E100, E360/E361, and D363/E364 in subdomain 1 of Dictyostelium actin were replaced with histidine residues by site-directed mutagenesis. Mutant actins were expressed in Dictyostelium cells and purified to homogeneity. The sliding movement of mutant actin filaments on heavy meromyosin attached to a glass surface was measured to assess the effect of the mutation on the motility of actin. For two C-terminal mutants, force generated by a single actin filament and myosin was also measured. These measurements indicated that both D24/D25 and E99/E100 are involved in ATP-driven sliding, whereas E360/E361/D363/E364 are not essential for ATP-driven sliding and force generation.

MeSH Terms
Actins/chemistry,genetics,metabolism Adenosine Triphosphate/metabolism Amino Acid Sequence Animals Dictyostelium/genetics,metabolism Histidine Models, Molecular Mutagenesis, Site-Directed Myosin Subfragments/metabolism Myosins/metabolism Protein Conformation Rabbits Recombinant Proteins/chemistry,metabolism
Chemicals
Actins Myosin Subfragments Recombinant Proteins Histidine Adenosine Triphosphate Myosins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Johara M
Department of Pure and Applied Sciences, College of Arts and Sciences, University of Tokyo, Japan.
Toyoshima Y Y
Ishijima A
Kojima H
Yanagida T
Sutoh K
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.90.6.2127
Published
1993-03-15
Pages
2127-31
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC46038
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.90.6.2127 PMC Full Text View journal details
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