Role of gizzard myosin light chains in calcium binding.
The contraction of molluscan and vertebrate smooth muscles is regulated by myosin. Although the myosin and its associated two subunits, the regulatory light chain and the essential light chain, constitute the Ca2+ regulatory system in both types of muscles, the mechanisms by which Ca2+ signal is transduced are quite different. In molluscan muscles, the direct binding of Ca2+ to the regulatory system triggers muscle contraction. In vertebrate smooth muscles, however, phosphorylation of the regulatory light chain is the major triggering mechanism. We measured Ca2+ binding in gizzard myosin and in hybrids of scallop myosin containing gizzard regulatory light chain or in hybrids of scallop regulatory domain containing gizzard essential light chain. Isolated chicken gizzard myosin did not bind Ca2+ in the range of pCa 8.0 to 5.0 in the presence of 2 mM MgCl2, supporting the lack of the specific Ca(2+)-binding site in gizzard myosin. Phosphorylation of the regulatory light chain did not generate a specific (Ca2+)-binding site. The hybrid scallop myosin containing gizzard regulatory light chain showed a similar Ca2+ binding as native scallop myosin with a one to one stoichiometry of Ca2+ to myosin head saturating at about pCa 6.0 at pH 7.6. In contrast, the hybrid scallop regulatory domain containing gizzard essential light chain did not bind Ca2+ either at pCa 6.0 or at pCa 8.0. Control preparations reconstituted with scallop essential light chains bound 0.69 mol per mol Ca2+ at pCa 6.0 with no binding at pCa 8.0.(ABSTRACT TRUNCATED AT 250 WORDS)
Kwon
H
H
Biophysics Programme, Brandeis University, Waltham, MA 02254.
Melandri
F D
FD
Szent-Györgyi
A G
AG
D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D002645:Chickens / Q000378:metabolism*; D005895:Gizzard, Avian; D008974:Mollusca; D009218:Myosins / Q000378:metabolism / Q000502:physiology*; D010766:Phosphorylation; D011485:Protein Binding; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction; D013045:Species Specificity
D009218:Myosins; D002118:Calcium