Conformation of the cytoplasmic domain of phospholamban by NMR and CD
- 1 January 1994
- journal article
- research article
- Published by Taylor & Francis in Molecular Membrane Biology
- Vol. 11 (4) , 263-269
- https://doi.org/10.3109/09687689409160436
Abstract
Nuclear magnetic resonance (NMR) and circular dichroism (CD) spectroscopy have been used to characterize the conformation of the putative cytoplasmic domain of phospholamban (PLB), an oligomeric membrane-bound protein which regulates the activity of the cardiac sarcoplasmic reticulum Ca(2+)-dependent ATPase. In aqueous solution the 25-residue peptide adopts a number of rapidly interconverting conformers with no secondary structural type obviously predominating. However, in trifluoroethanol (TFE) the conformation, while still highly dynamic, is characterized by a high proportion of helical structures. Evidence for this is provided by alpha CH chemical shifts and low NH chemical shift temperature coefficients, small NH-alpha CH intraresidue scalar coupling constants, a substantial number of distinctive interresidue nuclear Overhauser effects (NOEs) [dNN(i, i + 1), d alpha N(i, i + 3), d alpha beta(i, i + 3) and d alpha N(i, i + 4)] and characteristic CD bands at 190 (positive), 206 (negative) and 222 nm (negative). The helicity is interrupted around Pro-21. The activity of PLB is regulated by phosphorylation at either Ser-16 or Thr-17. CD shows that phosphorylation at Ser-16 by the cAMP-activated protein kinase causes about an 11% decrease in alpha-helical content in TFE.Keywords
This publication has 20 references indexed in Scilit:
- Helix geometry in proteinsPublished by Elsevier ,2004
- Interaction of calmodulin with phospholamban and caldesmon: comparative studies by 1H-NMR spectroscopyBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1992
- Evidence for a phosphorylation‐induced conformational change in phospholamban cytoplasmic domain by CD analysisFEBS Letters, 1992
- Regulation of the calcium ion pump of sarcoplasmic reticulum: Reversible inhibition by phospholamban and by the calmodulin binding domain of the plasma membrane calcium pumpBiochemistry, 1992
- Relationship between nuclear magnetic resonance chemical shift and protein secondary structureJournal of Molecular Biology, 1991
- Secondary structure of detergent-solubilized phospholamban, a phosphorylatable, oligomeric protein of cardiac sarcoplasmic reticulumBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1989
- Nuclear Overhauser effects in aqueous solution as dynamic probes in short linear peptidesFEBS Letters, 1988
- Improved spectral resolution in COSY 1H NMR spectra of proteins via double quantum filteringBiochemical and Biophysical Research Communications, 1983
- Application of phase sensitive two-dimensional correlated spectroscopy (COSY) for measurements of 1H-1H spin-spin coupling constants in proteinsBiochemical and Biophysical Research Communications, 1983
- Determination of the secondary structures of proteins by circular dichroism and optical rotatory dispersionBiochemistry, 1972