PMID: 3758070Sep 15, 1986Paper

Calcium release from calmodulin and its C-terminal or N-terminal halves in the presence of the calmodulin antagonists phenoxybenzamine and melittin measured by stopped-flow fluorescence with Quin 2 and intrinsic tyrosine. Inhibition of calmodulin-dependent protein kinase of cardiac sarcoplasmic reticulum

European Journal of Biochemistry
J SukoG Hellmann

Abstract

Calcium dissociation from the C-terminal and N-terminal halves of calmodulin, intact bovine brain calmodulin and the respective phenoxybenzamine complexes or melittin complexes was measured directly by stopped-flow fluorescence with the calcium chelator Quin 2 and, when possible, also by protein fluorescence using endogenous tyrosine fluorescence by mixing with EGTA. Calcium dissociation from the C-terminal half of calmodulin, which contains only the two high-affinity calcium-binding sites, and from intact calmodulin was monophasic, with good correlation of the rates of calcium dissociation obtained by the two methods. The apparent rates with Quin 2 and endogenous tyrosine fluorescence were 13.4 s-1 and 12.8 s-1, respectively, in the C-terminal half and 10.5 s-1 and 10.8 s-1, respectively, in intact calmodulin (pH 7.0, 25 degrees C, 100 mM KCl). Alkylation of the C-terminal half resulted in a biphasic calcium dissociation (Quin 2: kobs 1.90 s-1 and 0.73 s-1 respectively; tyrosine: kobs 1.65 s-1 and 0.61 s-1 respectively). Alkylation of intact calmodulin resulted in a four-phase calcium dissociation measured with Quin 2 (kobs 85.3 s-1, 11.1 s-1, 1.92 s-1 and 0.59 s-1); the latter two phases are assumed to represent calcium relea...Continue Reading

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Citations

Jan 1, 1989·Pharmacology & Therapeutics·M L VeiglW D Sedwick
Sep 7, 2000·Protein Science : a Publication of the Protein Society·L MasinoP M Bayley
Apr 15, 1987·European Journal of Biochemistry·M Zimmer, F Hofmann
Feb 16, 2002·European Journal of Biochemistry·Makoto TakanoTsuneyoshi Horigome
Jan 30, 2015·British Journal of Pharmacology·Doriano FabbroHenrik Moebitz
Feb 7, 1997·The Journal of Biological Chemistry·S E BrownP M Bayley
Jan 30, 2002·Biochemistry·Tomohiko Murase, Takayoshi Iio
Feb 8, 1988·Biochimica Et Biophysica Acta·W WyskovskyJ Suko
Mar 12, 2008·Biochimica Et Biophysica Acta·Liubov Lemberskiy-KuzinZvi Priel

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