Chimeric myosin regulatory light chains identify the subdomain responsible for regulatory function.
Regulatory light chains, located on the 'motor' head domains of myosin, belong to the family of Ca2+ binding proteins that consist of four 'EF-hand' subdomains. Vertebrate regulatory light chains can be divided into two functional classes: (i) in smooth/non-muscle myosins, phosphorylation of the light chains by a calcium/calmodulin-dependent kinase regulates both interaction of the myosin head with actin and assembly of the myosin into filaments, (ii) the light chains of skeletal muscle myosins are similarly phosphorylated, but they play no apparent role in regulation. To discover the basis for the difference in regulatory properties of these two classes of light chains, we have synthesized in Escherichia coli, chimeric mutants composed of subdomains derived from the regulatory light chains of chicken skeletal and smooth muscle myosins. The regulatory capability of these mutants was analysed by their ability to regulate molluscan myosin. Using this test system, we identified the third subdomain of the regulatory light chain as being responsible for controlling not only the actin-myosin interaction, but also myosin filament assembly.
Rowe
T
T
MRC Laboratory of Molecular Biology, Cambridge, UK.
Kendrick-Jones
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002645:Chickens; D004277:DNA, Single-Stranded; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D008974:Mollusca; D009132:Muscles / Q000201:enzymology / Q000378:metabolism; D009218:Myosins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D010957:Plasmids; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics; D016415:Sequence Alignment
D004277:DNA, Single-Stranded; D011993:Recombinant Fusion Proteins; D009218:Myosins; D002118:Calcium