Isolation of the regulatory domain of scallop myosin

Role of the essential light chain in calcium binding

Hyockman Kwon, Elizabeth B. Goodwin, L. Nyitray, Elise Berliner, Elizabeth O'Neall-Hennessey, Francesco D. Melandri, Andrew G. Szent-Györgyi

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

The regulatory domain of scallop myosin, consisting of a regulatory light chain (R-LC), an essential light chain (E-LC), and a portion of heavy chain, occupies the neck region of myosin. This domain is directly involved in the regulation of molluscan muscle contraction, which is triggered by direct Ca2+ binding to myosin. We have isolated a soluble functional complex (regulatory complex) comprised of R-LC, E-LC, and a 10-kDa heavy chain fragment in a 1:1:1 stoichiometry by clostripain digestion of the myosin head (papain subfragment 1). N termini of the heavy chain fragments were oither leucine-812 or valine-817. The isolated complex retained the specific Ca2+-binding site and bound Ca2+ with a similar affinity and selectivity as myosin. The individual components of the regulatory complex were isolated after complete denaturation with guanidine hydrochloride. The regulatory complex was reconstituted from isolated light chains and the heavy chain fragment. The renatured complex regained Ca2+ binding quantitatively. To elucidate the function of the E-LC in Ca2+ binding, we constructed hybrid regulatory complexes. The hybrid complexes reconstituted with molluscan E-LC and R-LC regained the specific Ca2+-binding site, whereas the hybrid complex formed with rabbit skeletal E-LC [alkali LC 2 (A2-LC)] and scallop R-LC did not. The results demonstrate that E-LCs from myosins regulated by direct Ca2+ binding are required for the specific Ca2+ binding in the molluscan muscle. (.

Original languageEnglish
Pages (from-to)4771-4775
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number12
Publication statusPublished - 1990

Fingerprint

Pectinidae
Myosin Light Chains
Calcium
Light
Myosins
clostripain
Binding Sites
Papain
Guanidine
Valine
Alkalies
Muscle Contraction
varespladib methyl
Leucine
Digestion
Neck

Keywords

  • Calcium regulation
  • Muscle
  • Renaturation

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Kwon, H., Goodwin, E. B., Nyitray, L., Berliner, E., O'Neall-Hennessey, E., Melandri, F. D., & Szent-Györgyi, A. G. (1990). Isolation of the regulatory domain of scallop myosin: Role of the essential light chain in calcium binding. Proceedings of the National Academy of Sciences of the United States of America, 87(12), 4771-4775.

Isolation of the regulatory domain of scallop myosin : Role of the essential light chain in calcium binding. / Kwon, Hyockman; Goodwin, Elizabeth B.; Nyitray, L.; Berliner, Elise; O'Neall-Hennessey, Elizabeth; Melandri, Francesco D.; Szent-Györgyi, Andrew G.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 12, 1990, p. 4771-4775.

Research output: Contribution to journalArticle

Kwon, H, Goodwin, EB, Nyitray, L, Berliner, E, O'Neall-Hennessey, E, Melandri, FD & Szent-Györgyi, AG 1990, 'Isolation of the regulatory domain of scallop myosin: Role of the essential light chain in calcium binding', Proceedings of the National Academy of Sciences of the United States of America, vol. 87, no. 12, pp. 4771-4775.
Kwon, Hyockman ; Goodwin, Elizabeth B. ; Nyitray, L. ; Berliner, Elise ; O'Neall-Hennessey, Elizabeth ; Melandri, Francesco D. ; Szent-Györgyi, Andrew G. / Isolation of the regulatory domain of scallop myosin : Role of the essential light chain in calcium binding. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 12. pp. 4771-4775.
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