Biological Activities of the Peptides Obtained by Digestion of Troponin C and Calmodulin with Thrombin

Overview

Journal Biochem J

Specialty Biochemistry

Date 1981 Apr 1

PMID 6895466

Citations 4

Authors

C M Wall

R J Grand

S V PERRY

Affiliations

Soon will be listed here.

Abstract

1. Troponin C and calmodulin were not digested by thrombin at a significant rate in the presence of Ca2+. 2. In the presence of EGTA, troponin C was digested by thrombin to yield three peptides, TH1 (residues 1--120), TH3 (residues 1--100) and TH2 (residues 121--159). 3. In the presence of EGTA calmodulin was digested by thrombin giving two peptides, TM1 (residues 1--106) and TM2 (residues 107--148). 4. The electrophoretic mobilities of peptides TH1 and TM1 were increased at pH 8.6 by Ca2+ both in the presence and absence of urea. The mobilities of peptides TH2 and TM2 were unaltered under these conditions. 5. Peptides TH1, TH2 and tM1 formed complexes with troponin I on polyacrylamide gels at pH 8.6 in the presence of Ca2+. 6. The phosphorylation of troponin I by cyclic AMP-dependent protein kinase was significantly inhibited by peptides TH1 and TH3 and to a lesser extent by peptide TM1. 7. The calmodulin peptide TM1 activated myosin light-chain kinase when present in large molar excess. Peptide TM2 did not activate the enzyme.

Citing Articles

Peptide and metal ion-dependent association of isolated helix-loop-helix calcium binding domains: studies of thrombic fragments of calmodulin.

Brokx R, Vogel H Protein Sci. 2000; 9(5):964-75.

PMID: 10850806 PMC: 2144632. DOI: 10.1110/ps.9.5.964.

Phosphorylation of calmodulin on Tyr99 selectively attenuates the action of calmodulin antagonists on type-I cyclic nucleotide phosphodiesterase activity.

Saville M, Houslay M Biochem J. 1994; 299 ( Pt 3):863-8.

PMID: 8192677 PMC: 1138100. DOI: 10.1042/bj2990863.

Calmodulin and the regulation of smooth muscle contraction.

Walsh M Mol Cell Biochem. 1994; 135(1):21-41.

PMID: 7816054 DOI: 10.1007/BF00925958.

Substitution at position 116 of Schizosaccharomyces pombe calmodulin decreases its stability under nitrogen starvation and results in a sporulation-deficient phenotype.

Takeda T, Imai Y, Yamamoto M Proc Natl Acad Sci U S A. 1989; 86(24):9737-41.

PMID: 2690071 PMC: 298576. DOI: 10.1073/pnas.86.24.9737.

References

Weber K, Osborn M . The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969; 244(16):4406-12. View

SANGER F, Thompson E . Halogenation of tyrosine during acid hydrolysis. Biochim Biophys Acta. 1963; 71:468-71. DOI: 10.1016/0006-3002(63)91108-9. View

Gray W, Smith J . Rapid sequence analysis of small peptides. Anal Biochem. 1970; 33(1):36-42. DOI: 10.1016/0003-2697(70)90436-7. View

Hartley B . Strategy and tactics in protein chemistry. Biochem J. 1970; 119(5):805-22. PMC: 1179482. DOI: 10.1042/bj1190805f. View

Ebashi S, Wakabayashi T, EBASHI F . Troponin and its components. J Biochem. 1971; 69(2):441-5. DOI: 10.1093/oxfordjournals.jbchem.a129486. View

Swank R, MUNKRES K . Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971; 39(2):462-77. DOI: 10.1016/0003-2697(71)90436-2. View

Wilkinson J, PERRY S, Cole H, Trayer I . The regulatory proteins of the myofibril. Separation and biological activity of the components of inhibitory-factor preparations. Biochem J. 1972; 127(1):215-28. PMC: 1178576. DOI: 10.1042/bj1270215. View

Gould J, Cather R, Winget G . Advantages of the use of Cerenkov vounting for determination of P 32 in photophosphorylation research. Anal Biochem. 1972; 50(2):540-8. DOI: 10.1016/0003-2697(72)90064-4. View

COLLINS J, Potter J, Horn M, Wilshire G, JACKMAN N . The amino acid sequence of rabbit skeletal muscle troponin C: gene replication and homology with calcium-binding proteins from carp and hake muscle. FEBS Lett. 1973; 36(3):268-72. DOI: 10.1016/0014-5793(73)80388-6. View

10.

Head J, PERRY S . The interaction of the calcium-binding protein (troponin C) with bivalent cations and the inhibitory protein (troponin I). Biochem J. 1974; 137(2):145-54. PMC: 1166100. DOI: 10.1042/bj1370145. View

11.

Potter J, Gergely J . The calcium and magnesium binding sites on troponin and their role in the regulation of myofibrillar adenosine triphosphatase. J Biol Chem. 1975; 250(12):4628-33. View

12.

PERRY S, Cole H . Phosphorylation of troponin and the effects of interactions between the components of the complex. Biochem J. 1974; 141(3):733-43. PMC: 1168180. DOI: 10.1042/bj1410733. View

13.

Kretsinger R, Barry C . The predicted structure of the calcium-binding component of troponin. Biochim Biophys Acta. 1975; 405(1):40-52. DOI: 10.1016/0005-2795(75)90312-8. View

14.

Cole H, PERRY S . The phosphorylation of troponin I from cardiac muscle. Biochem J. 1975; 149(3):525-33. PMC: 1165658. DOI: 10.1042/bj1490525. View

15.

Jackson P, Amphlett G, PERRY S . The primary structure of troponin T and the interaction with tropomyosin. Biochem J. 1975; 151(1):85-97. PMC: 1172328. DOI: 10.1042/bj1510085. View

16.

COLLINS J . Homology of myosin DTNB light chain with alkali light chains, troponin C and parvalbumin. Nature. 1976; 259(5545):699-700. DOI: 10.1038/259699a0. View

17.

Syska H, Wilkinson J, Grand R, PERRY S . The relationship between biological activity and primary structure of troponin I from white skeletal muscle of the rabbit. Biochem J. 1976; 153(2):375-87. PMC: 1172583. DOI: 10.1042/bj1530375. View

18.

DRABIKOWSKI W, Kuznicki J, Grabarek Z . Similarity in Ca2+-induced changes between troponic-C and protein activator of 3':5'-cyclic nucleotide phosphodiesterase and their tryptic fragments. Biochim Biophys Acta. 1977; 485(1):124-33. DOI: 10.1016/0005-2744(77)90199-1. View

19.

Weeks R, PERRY S . A region of the troponic C molecule involved in interaction with troponin I [proceedings]. Biochem Soc Trans. 1977; 5(5):1391-2. DOI: 10.1042/bst0051391. View

20.

Pires E, PERRY S . Purification and properties of myosin light-chain kinase from fast skeletal muscle. Biochem J. 1977; 167(1):137-46. PMC: 1183630. DOI: 10.1042/bj1670137. View