caa a22 4500 606242783 CHVBK 20210128101320.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s12551-014-0157-z doi (NATIONALLICENCE)springer-10.1007/s12551-014-0157-z Phospholamban phosphorylation, mutation, and structural dynamics: a biophysical approach to understanding and treating cardiomyopathy [Elektronische Daten] [Naa-Adjeley Ablorh, David Thomas] We review the recent development of novel biochemical and spectroscopic methods to determine the site-specific phosphorylation, expression, mutation, and structural dynamics of phospholamban (PLB), in relation to its function (inhibition of the cardiac calcium pump, SERCA2a), with specific focus on cardiac physiology, pathology, and therapy. In the cardiomyocyte, SERCA2a actively transports Ca2+ into the sarcoplasmic reticulum (SR) during relaxation (diastole) to create the concentration gradient that drives the passive efflux of Ca2+ required for cardiac contraction (systole). Unphosphorylated PLB (U-PLB) inhibits SERCA2a, but phosphorylation at S16 and/or T17 (producing P-PLB) changes the structure of PLB to relieve SERCA2a inhibition. Because insufficient SERCA2a activity is a hallmark of heart failure, SERCA2a activation, by gene therapy (Andino et al. 2008; Fish et al. 2013; Hoshijima et al. 2002; Jessup et al. 2011) or drug therapy (Ferrandi et al. 2013; Huang 2013; Khan et al. 2009; Rocchetti et al. 2008; Zhang et al. 2012), is a widely sought goal for treatment of heart failure. This review describes rational approaches to this goal. Novel biophysical assays, using site-directed labeling and high-resolution spectroscopy, have been developed to resolve the structural states of SERCA2a-PLB complexes in vitro and in living cells. Novel biochemical assays, using synthetic standards and multidimensional immunofluorescence, have been developed to quantitate PLB expression and phosphorylation states in cells and human tissues. The biochemical and biophysical properties of U-PLB, P-PLB, and mutant PLB will ultimately resolve the mechanisms of loss of inhibition and gain of inhibition to guide therapeutic development. These assays will be powerful tools for investigating human tissue samples from the Sydney Heart Bank, for the purpose of analyzing and diagnosing specific disorders. International Union for Pure and Applied Biophysics (IUPAB) and Springer-Verlag Berlin Heidelberg, 2015 Phospholamban nationallicence SERCA2a nationallicence Phosphorylation nationallicence Subunit model nationallicence Loss-of-inhibition mutants nationallicence Ablorh Naa-Adjeley Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, 55455, Minneapolis, MN, USA aut Thomas David Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, 55455, Minneapolis, MN, USA aut Biophysical Reviews Springer Berlin Heidelberg 7/1(2015-03-01), 63-76 1867-2450 7:1<63 2015 7 12551 https://doi.org/10.1007/s12551-014-0157-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s12551-014-0157-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ablorh Naa-Adjeley Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, 55455, Minneapolis, MN, USA aut NATIONALLICENCE 700 1- Thomas David Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, 55455, Minneapolis, MN, USA aut NATIONALLICENCE 773 0- Biophysical Reviews Springer Berlin Heidelberg 7/1(2015-03-01), 63-76 1867-2450 7:1<63 2015 7 12551