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PMID: 11287639 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Three-dimensional image reconstruction of dephosphorylated smooth muscle heavy meromyosin reveals asymmetry in the interaction between myosin heads and placement of subfragment 2.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 98 ·No. 8 ·2001-04-10 ·Pages 4361-6

Wendt T, Taylor D, Trybus KM, Taylor K

Abstract

Regulation of the actin-activated ATPase of smooth muscle myosin II is known to involve an interaction between the two heads that is controlled by phosphorylation of the regulatory light chain. However, the three-dimensional structure of this inactivated form has been unknown. We have used a lipid monolayer to obtain two-dimensional crystalline arrays of the unphosphorylated inactive form of smooth muscle heavy meromyosin suitable for structural studies by electron cryomicroscopy of unstained, frozen-hydrated specimens. The three-dimensional structure reveals an asymmetric interaction between the two myosin heads. The ATPase activity of one head is sterically "blocked" because part of its actin-binding interface is positioned onto the converter domain of the second head. ATPase activity of the second head, which can bind actin, appears to be inhibited through stabilization of converter domain movements needed to release phosphate and achieve strong actin binding. When the subfragment 2 domain of heavy meromyosin is oriented as it would be in an actomyosin filament lattice, the position of the heads is very different from that needed to bind actin, suggesting an additional contribution to ATPase inhibition in situ.

MeSH Terms
Animals Chickens Microscopy, Electron Models, Molecular Muscle, Smooth/chemistry Myosin Subfragments/chemistry,ultrastructure Phosphorylation Protein Conformation
Chemicals
Myosin Subfragments
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wendt T
Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306, USA.
Taylor D
Trybus K M
Taylor 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.071051098
Published
2001-04-10
Epub
2001-00-03
Pages
4361-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC31840
Subset
IM
Grants
NHLBI NIH HHS · R01 HL038113 · United States
Databases
PDB
1I84
External Links
PubMed source DOI: 10.1073/pnas.071051098 PMC Full Text View journal details
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