Interaction of calmodulin with skeletal muscle myosin light chain kinase.

Studies on myosin light chain kinase isolated from rabbit skeletal muscle show that the enzyme has a molecular weight of 80,000--84,000 with a sedimentation coefficient of 3.2 S and an apparent Stokes radius of 53 A. Gel filtration chromatography with a 3H-labeled calmodulin using a Hummel--Dryer technique shows that the enzyme will bind 1 mol of calmodulin per mol of enzyme, with an affinity of (1.9 +/- 0.5) x 10(7) M-1 in the absence of substrate. The calmodulin dependence of enzyme activation at limiting Mg2+ and light chain concentrations confirms this observation. The calcium dependence of activation of the enzyme--calmodulin complex is characterized by a Hill coefficient of 2.5, with half-activation occurring at 6.6 x 10(-7) M Ca2+. The amino acid composition shows a high percentage (9.1%) of proline, which may account for the large apparent Stokes radius and no clear resemblance to other skeletal muscle proteins. A comparison of the amino acid composition with that from turkey gizzard shows some resemblance.

[1]  D. Blumenthal,et al.  Activation of skeletal muscle myosin light chain kinase by calcium(2+) and calmodulin. , 1980, Biochemistry.

[2]  C. Klee,et al.  Positive cooperative binding of calcium to bovine brain calmodulin. , 1980, Biochemistry.

[3]  C. L. Johnson,et al.  Ca2+-dependent regulation of guinea pig brain adenylate cyclase. , 1980, The Journal of biological chemistry.

[4]  J. Cavadore,et al.  Homologous calcium-binding proteins in the activation of skeletal, cardiac, and smooth muscle myosin light chain kinases. , 1980, The Journal of biological chemistry.

[5]  T. H. Crouch,et al.  Magnetic osmometry: association of two peptic fragments from bovine serum albumin at micromolar concentrations. , 1980, Biochemistry.

[6]  W. Y. Cheung,et al.  Calmodulin plays a pivotal role in cellular regulation. , 1980, Science.

[7]  D. A. Small,et al.  Isolation of cardiac myofibrils and myosin light chains with in vivo levels of light chain phosphorylation. , 1979, Biochimica et biophysica acta.

[8]  L. K. Lane,et al.  Reaction of purified (Na,K)-ATPase with the fluorescent sulfhydryl probe 2-(4'-maleimidylanilino)naphthalene 6-sulfonic acid. Characterization and the effects of ligands. , 1979, The Journal of biological chemistry.

[9]  A. Nairn,et al.  The role of calmodulin in the myosin light-chain kinase system [proceedings]. , 1979, Biochemical Society transactions.

[10]  J. Stull,et al.  Myosin light chain phosphorylation and phosphorylase A activity in rat extensor digitorum longus muscle. , 1979, Biochemical and biophysical research communications.

[11]  M. Bárány,et al.  Phosphorylation-dephosphorylation of the 18,000-dalton light chain of myosin during the contraction-relaxation cycle of frog muscle. , 1979, The Journal of biological chemistry.

[12]  A. Nairn,et al.  Calmodulin and myosin light-chain kinase of rabbit fast skeletal muscle. , 1979, The Biochemical journal.

[13]  Davenport Hw.  Some notes on preparing a history of a department of physiology. , 1979 .

[14]  K. Yagi,et al.  Purification of modulator-deficient myosin light-chain kinase by modulator protein-Sepharose affinity chromatography. , 1978, Journal of biochemistry.

[15]  P. Cohen,et al.  Identification of the Ca2+‐dependent modulator protein as the fourth subunit of rabbit skeletal muscle phosphorylase kinase , 1978, FEBS letters.

[16]  K. Yagi,et al.  Identification of an activator protein for myosin light chain kinase as the Ca2+-dependent modulator protein. , 1978, The Journal of biological chemistry.

[17]  L. Smillie,et al.  The amino acid sequence of rabbit skeletal alpha-tropomyosin. The NH2-terminal half and complete sequence. , 1978, The Journal of biological chemistry.

[18]  A. Means,et al.  Physicochemical properties of rat testis Ca2+-dependent regulator protein of cyclic nucleotide phosphodiesterase. Relationship of Ca2+-binding, conformational changes, and phosphodiesterase activity. , 1977, The Journal of biological chemistry.

[19]  S. Perry,et al.  Purification and properties of myosin light-chain kinase from fast skeletal muscle. , 1977, The Biochemical journal.

[20]  P. Fasella,et al.  An assessment of some of the methods available for the determination of molecular weights of proteins as applied to aspartate aminotransferase from pig heart. , 1976, European journal of biochemistry.

[21]  A. Moir,et al.  Phosphorylation of troponin I and the inotropic effect of adrenaline in the perfused rabbit heart , 1976, Nature.

[22]  L. Smillie,et al.  Amino-acid sequence of tropomyosin-binding component of rabbit skeletal muscle troponin. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[23]  J. H. Collins,et al.  The primary structure of actin from rabbit skeletal muscle. Completion and analysis of the amino acid sequence. , 1975, The Journal of biological chemistry.

[24]  J. Wilkinson,et al.  The amino acid sequence of troponin I from rabbit skeletal muscle. , 1975, The Biochemical journal.

[25]  B. Vanyushin,et al.  Granulated hydroxyapatite: preparation and chromatographic properties. , 1974, Analytical biochemistry.

[26]  C. Tanford,et al.  Molecular characterization of proteins in detergent solutions. , 1974, Biochemistry.

[27]  G. Frank,et al.  The amino-acid sequence of the alkali light chains of rabbit skeletal-muscle myosin. , 1974, European journal of biochemistry.

[28]  J. Weltman,et al.  Relatedness among contractile and membrane proteins: evidence for evolution from common ancestral genes. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[29]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[30]  S. Edelstein,et al.  Ultracentrifuge studies with absorption optics. IV. Molecular weight determinations at the microgram level. , 1966, Biochemistry.

[31]  D. A. Yphantis.  EQUILIBRIUM ULTRACENTRIFUGATION OF DILUTE SOLUTIONS. , 1964, Biochemistry.

[32]  W. F. Harrington,et al.  A model for the myosin molecule. , 1960, Biochimica et biophysica acta.

[33]  Oliver H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[34]  T. Mcmeekin,et al.  Apparent Specific Volume of α-Casein and β-Casein and the Relationship of Specific Volume to Amino Acid Composition , 1949 .

[35]  I. Kuntz,et al.  The properties of water in biological systems. , 1974, Annual review of biophysics and bioengineering.

[36]  S. Lowey,et al.  An Immunochemical Approach to the Interaction of Light and Heavy Chains in Myosin , 1973 .

[37]  D. Kupke.  Density and Volume Change Measurements , 1973 .

[38]  G. K. Ackers.  Studies of protein ligand binding by gel permeation techniques. , 1973, Methods in enzymology.

[39]  S. Leach.  Physical principles and techniques of protein chemistry , 1969 .