Conformational transition accompanying the binding of Ca2+ to the protein activator of 3',5'-cyclic adenosine monophosphate phosphodiesterase.

作者: C. B. Klee

DOI: 10.1021/BI00624A033

关键词:

摘要: The Ca2+-dependent protein activator of 3':5'-cyclic adenosine monophosphate phosphodiesterase is shown to undergo a conformational transition upon binding 2 mol Ca2+/mol activator. Circular dichroic studies indicate that Ca2+ induces an increase 5-8% in alpha-helix content with concomitant decrease the amount random coil. In absence and presence [ethylenebis(oxoethylenenitrilo)]tetraacetic acid (EGTA), contains 30-35% alpha helix, 50% coil, 15-20% beta-pleated sheat. Spectrophotometric titration indicates two tyrosyl residues have pK's 10.4 11.9 are therefore different environments. Ca2+-induced change accompanied by increased exposure protons partially exposed tyrosine, as shift its pK from 10.). Increased solvation also consistent negative difference spectrum at 287 279 nm seen binding. Modification environment all or some phenylalanine part accompanying A new rapid purification procedure which yields large amounts (25-30% yields) homogenous direct sensitive assay for cAMP described.

参考文章(38)
Charles C. Bigelow, Ultraviolet Spectral Studies of Acid- and Base-denatured Ribonuclease Journal of Biological Chemistry. ,vol. 236, pp. 1706- 1710 ,(1961) , 10.1016/S0021-9258(19)63289-8
Yasuhiko Nozaki, Charles Tanford, [84] Examination of titration behavior Methods in Enzymology. ,vol. 11, pp. 715- 734 ,(1967) , 10.1016/S0076-6879(67)11088-4
Östen Levin, [2] Column chromatography of proteins: Calcium phosphate Methods in Enzymology. ,vol. 5, pp. 27- 32 ,(1962) , 10.1016/S0076-6879(62)05184-8
G. Schultz, E. Böhme, J.G. Hardman, [2] Separation and purification of cyclic nucleotides by ion-exchange resin column chromatography Hormone Action Part C: Cyclic Nucleotides. ,vol. 38, pp. 9- 20 ,(1974) , 10.1016/0076-6879(74)38005-6
T.T. Herskovits, [86] Difference spectroscopy Methods in Enzymology. ,vol. 11, pp. 748- 775 ,(1967) , 10.1016/S0076-6879(67)11090-2
C.H.W. Hirs, [19] Performic acid oxidation Methods in Enzymology. ,vol. 11, pp. 197- 199 ,(1967) , 10.1016/S0076-6879(67)11021-5
Ying Ming Lin, Yung Pin Liu, Wai Yiu Cheung, [39] Purification and characterization of a protein activator of cyclic nucleotide phosphodiesterase from bovine brain☆ Methods in Enzymology. ,vol. 38, pp. 262- 273 ,(1974) , 10.1016/0076-6879(74)38042-1
Marion L. Greaser, John Gergely, Purification and Properties of the Components from Troponin Journal of Biological Chemistry. ,vol. 248, pp. 2125- 2133 ,(1973) , 10.1016/S0021-9258(19)44195-1
Ying Ming Lin, Yung Pin Liu, Wai Yiu Cheung, Cyclic 3' : 5'-Nucleotide Phosphodiesterase PURIFICATION, CHARACTERIZATION, AND ACTIVE FORM OF THE PROTEIN ACTIVATOR FROM BOVINE BRAIN Journal of Biological Chemistry. ,vol. 249, pp. 4943- 4954 ,(1974) , 10.1016/S0021-9258(19)42412-5