H-1- and H-2-NMR studies of a fragment of PMP1, a regulatory subunit associated with the yeast plasma membrane H+-ATPase. Conformational properties and lipid-peptide interactions

PMP1 is a 38-residue polypeptide associated with the yeast plasma membrane H+-ATPase, found to regulate the enzyme activity. To investigate the molecular basis of the PMP1 biological function, the conformational properties of a synthetic PMP1 fragment, A18-F38, comprising the predicted C-terminal cytoplasmic domain and a part of the transmembrane anchor have been studied by H-1- and H-2-NMR spectroscopies. High resolution H-1-NMR experiments showed that, in deuterated DPC micelles, the A18-G34 segment adopts a well defined helix conformation. Our data suggest that the whole PMP1 molecule forms a unique helix whose axis might be slightly tilted with respect to the bilayer normal. Protonated DPC, DMPC and DMPS were incorporated in deuterated micelles containing the PMP1 fragment for studying Lipid-peptide interactions. Unusually strong and selective intermolecular NOEs between Lipid chain and peptide side chain protons, especially those of the unique Trp residue, were observed. Solid state H-2-NMR experiments performed on pure deuterated POPC and mixed deuterated POPC:POPS (5:1) bilayers revealed that the PMP1 fragment specifically interacts with negatively charged PS lipids ((C) Societe francaise de biochimie et biologie moleculaire / Elsevier, Paris)