Role of nucleotides in stabilization of the phospholamban/cardiac Ca(2+) pump inhibitory complex examined with use of metal fluorides

The FEBS Journal
Zhenhui Chen

Abstract

Phospholamban (PLB) inhibits the activity of the cardiac calcium pump SERCA2a. We previously showed that PLB with engineered Cys residues only cross-linked with the Ca(2+) -free E2 intermediate of SERCA2a. Formation of E2•PLB prevents Ca(2+) binding at the high-affinity Ca(2+) binding sites, blocking the enzyme kinetic cycle. Here we further studied the synergistic action of PLB and ATP on E2 in terms of prevention of formation of the phosphorylated E2P-like states stabilized by metal fluorides. SERCA2a was co-expressed in insect cell microsomes with PLB mutants of normal or super-inhibitory strength, with cross-linkable mutations at either the cytosolic side (N30C) or the luminal side (V49C) of PLB. For normal-strength PLB mutants, in the absence of nucleotide, metal fluorides totally inhibited both SERCA2a enzyme activity and cross-linking of PLB to SERCA2a at both sites, suggesting that PLB dissociates from SERCA2a in the E2P-like states. However, under the same conditions, super-inhibitory PLB mutants prevented total enzyme inhibition by metal fluorides. Further, the cross-linking of super-inhibitory PLB to SERCA2a was only partially inhibited by metal fluorides, but was drastically restored upon sequential addition of ATP....Continue Reading

References

Oct 28, 1998·Physiological Reviews·H K Simmerman, L R Jones
Jan 15, 2003·Proceedings of the National Academy of Sciences of the United States of America·Chikashi ToyoshimaDavid H MacLennan
Feb 25, 2003·Annual Review of Biophysics and Biomolecular Structure·David L Stokes, N Michael Green
Apr 1, 2006·Journal of Molecular Biology·Christine B KarimDavid D Thomas
May 20, 2006·The EMBO Journal·Anne-Marie Lund JensenPoul Nissen
Jan 31, 2007·Biochemistry·Jason R WaggonerJames E Mahaney
May 6, 2008·Archives of Biochemistry and Biophysics·Chikashi Toyoshima
Feb 4, 2011·The Journal of Biological Chemistry·Johannes D ClausenJens Peter Andersen
Jun 9, 2012·Circulation Research·Evangelia G Kranias, Roger J Hajjar
Feb 13, 2013·The Journal of Biological Chemistry·Maike BublitzPoul Nissen
Mar 5, 2013·Nature·Anne-Marie L WintherMorten J Buch-Pedersen
Sep 3, 2013·The Journal of Biological Chemistry·Brandy L AkinLarry R Jones

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Citations

Feb 23, 2021·Frontiers in Molecular Biosciences·Thomas BarbotLiliane Mouawad
May 1, 2020·Journal of Molecular and Cellular Cardiology·Wenbo HeZhenhui Chen

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