DOI: 10.1083/jcb.200105079. Myosin light chain kinase binding to a unique site on F-actin revealed by three-dimensional image reconstruction

Myosin light chain kinase binding to a unique site on F-actin revealed by three-dimensional image reconstruction

10.1083/jcb.200105079

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

Ca2+–calmodulin-dependent phosphorylation of myosin regulatory light chains by the catalytic COOH-terminal half of myosin light chain kinase (MLCK) activates myosin II in smooth and nonmuscle cells. In addition, MLCK binds to thin filaments in situ and F-actin in vitro via a specific repeat motif in its NH2 terminus at a stoichiometry of one MLCK per three actin monomers. We have investigated the structural basis of MLCK–actin interactions by negative staining and helical reconstruction. F-actin was decorated with a peptide containing the NH2-terminal 147 residues of MLCK (MLCK-147) that binds to F-actin with high affinity. MLCK-147 caused formation of F-actin rafts, and single filaments within rafts were used for structural analysis. Three-dimensional reconstructions showed MLCK density on the extreme periphery of subdomain-1 of each actin monomer forming a bridge to the periphery of subdomain-4 of the azimuthally adjacent actin. Fitting the reconstruction to the atomic model of F-actin revealed interaction of MLCK-147 close to the COOH terminus of the first actin and near residues 228–232 of the second. This unique location enables MLCK to bind to actin without interfering with the binding of any other key actin-binding proteins, including myosin, tropomyosin, caldesmon, and calponin.

Bibliography

Hatch, V., Zhi, G., Smith, L., Stull, J. T., Craig, R., & Lehman, W. (2001). Myosin light chain kinase binding to a unique site on F-actin revealed by three-dimensional image reconstruction. The Journal of Cell Biology, 154(3), 611â618.

Authors 6

Victoria Hatch (first)
Gang Zhi (additional)
Lula Smith (additional)
James T. Stull (additional)
Roger Craig (additional)
William Lehman (additional)

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 12:46 p.m.)
Deposited	2 years, 1 month ago (July 22, 2023, 7:46 a.m.)
Indexed	1 year, 1 month ago (July 21, 2024, 7:55 p.m.)
Issued	24 years, 1 month ago (July 30, 2001)
Published	24 years, 1 month ago (July 30, 2001)
Published Online	24 years, 1 month ago (July 30, 2001)
Published Print	24 years ago (Aug. 6, 2001)

Funders 0

None

BibTeX

@article{Hatch_2001, title={Myosin light chain kinase binding to a unique site on F-actin revealed by three-dimensional image reconstruction}, volume={154}, ISSN={0021-9525}, url={http://dx.doi.org/10.1083/jcb.200105079}, DOI={10.1083/jcb.200105079}, number={3}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Hatch, Victoria and Zhi, Gang and Smith, Lula and Stull, James T. and Craig, Roger and Lehman, William}, year={2001}, month=jul, pages={611–618} }

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