Conformation of the N-terminal segment of a monocysteine mutant of troponin I from cardiac muscle

Biochemistry
W J DongH C Cheung

Abstract

A monocysteine mutant of cardiac muscle troponin I, cTnI(S5C/C81I/C98S), was generated from a mouse cTnI cDNA clone and expressed in a bacterial system. Cys-5 was modified with the fluorescent sulfhydryl reagent IAANS to probe the conformation of the N-terminal extension of the mutant and the mutant complexed with cardiac muscle troponin C. Our emphasis was on the effect of phosphorylation of Ser-23 and Ser-24 by protein kinase A on the conformation of the N-terminal segment. Phosphorylation resulted in an 8-nm red-shift of the emission spectrum of the attached IAANS probe and a reduction of its quantum yield by a factor of 4-5. The intensity decay of nonphosphorylated IAANS-labeled mutant was complex and had to be described by a sum of three exponential terms, with lifetimes in the range 0.1-5 ns. A fourth component in the range 7-9 ns was required to describe the intensity decay of the phosphorylated mutant. Phosphorylation also reduced the weighted mean lifetime, consistent with the changes observed in the steady-state fluorescence parameters and a 33% decrease in the global rotational correlation time calculated from anisotropy decay data. This change in correlation time suggested a decrease in the axial ratio of the protei...Continue Reading

References

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Citations

Sep 12, 2003·Journal of Molecular and Cellular Cardiology·Paul R Rosevear, Natosha Finley
Jun 3, 1999·Biochimica Et Biophysica Acta·P DobryszyckiM Kochman
Dec 31, 2011·The Journal of Biological Chemistry·Zhiqun ZhouWen-Ji Dong
Mar 15, 2000·Protein Science : a Publication of the Protein Society·W J DongH C Cheung
Dec 6, 2011·American Journal of Physiology. Heart and Circulatory Physiology·Shirin AkhterJ-P Jin
Sep 20, 2002·Proceedings of the National Academy of Sciences of the United States of America·Louise J BrownPiotr G Fajer
Dec 20, 2000·European Journal of Biochemistry·D G WardI P Trayer
Sep 17, 2015·Chemical Biology & Drug Design·William SchlechtWenji Dong
Jun 1, 2008·Proteomics. Clinical Applications·Wenhai JinAnne M Murphy
Mar 2, 2005·Progress in Cardiovascular Diseases·Jason L McDonough, Jennifer E Van Eyk
Sep 12, 2006·Archives of Biochemistry and Biophysics·Wen-Ji DongHerbert C Cheung
Aug 14, 1999·The Journal of Biological Chemistry·Q Kleerekoper, J A Putkey
May 19, 2018·American Journal of Physiology. Heart and Circulatory Physiology·Victoria N ParikhEuan A Ashley
Jan 4, 2003·The Journal of Biological Chemistry·Wen-Ji DongHerbert C Cheung
Oct 21, 2020·Journal of Molecular and Cellular Cardiology·Ehsan KachooeiLouise J Brown

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