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PMID: 7858132 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Structural connectivity in actin: effect of C-terminal modifications on the properties of actin.

Biophysical journal ·Vol. 67 ·No. 5 ·1994-11-00 ·Pages 1957-64

Crosbie RH, Miller C, Cheung P, Goodnight T, Muhlrad A, Reisler E

Abstract

In this study, we use fluorescent probes and proteolytic digestions to demonstrate structural coupling between distant regions of actin. We show that modifications of Cys-374 in the C-terminus of actin slow the rate of nucleotide exchange in the nucleotide cleft. Conformational coupling between the C-terminus and the DNasal loop in subdomain II is observed in proteolytic digestion experiments in which a new C-terminal cleavage site is exposed upon DNasel binding. The functional consequences of C-terminal modification are evident from S-1 ATPase activity and the in vitro motility experiments with modified actins. Pyrene actin, labeled at Cys-374, activates S-1 ATPase activity only half as well as control actin. This reduction is attributed to a lower Vmax value because the affinity of pyrene actin to S-1 is not significantly altered. The in vitro sliding velocity of pyrene actin is also decreased. However, IAEDANS labeling of actin (also at Cys-374) enhances the Vmax of acto-S-1 ATPase activity and the in vitro sliding velocity by approximately 25%. These results are discussed in terms of conformational coupling between distant regions in actin and the functional implications of the interactions of actin-binding proteins with the C-terminus of actin.

MeSH Terms
Actins/chemistry,metabolism Animals Binding Sites Biophysical Phenomena Biophysics Fluorescent Dyes In Vitro Techniques Molecular Structure Myosins/metabolism Naphthalenesulfonates Peptide Fragments/chemistry,metabolism Polymers/chemistry,metabolism Protein Conformation Rabbits Structure-Activity Relationship
Chemicals
Actins Fluorescent Dyes Naphthalenesulfonates Peptide Fragments Polymers 1,5-I-AEDANS Myosins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Crosbie R H
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
Miller C
Cheung P
Goodnight T
Muhlrad A
Reisler E
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45 references, click to expand
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(94)80678-2
Published
1994-11-00
Pages
1957-64
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1225570
Subset
IM
Grants
NIAMS NIH HHS · R01 AR022031 · United States
NIAMS NIH HHS · AR22031 · United States
NIGMS NIH HHS · GM 07185 · United States
External Links
PubMed source DOI: 10.1016/S0006-3495(94)80678-2 PMC Full Text View journal details
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