PMP1 18-38, a yeast plasma membrane protein fragment, binds phosphatidylserine from bilayer mixtures with phosphatidylcholine: a (2)H-NMR study.
PMP1 is a 38-residue plasma membrane protein of the yeast Saccharomyces cerevisiae that regulates the activity of the H(+)-ATPase. The cytoplasmic domain conformation results in a specific interfacial distribution of five basic side chains, thought to strongly interact with anionic phospholipids. We have used the PMP1 18-38 fragment to carry out a deuterium nuclear magnetic resonance ((2)H-NMR) study for investigating the interactions between the PMP1 cytoplasmic domain and phosphatidylserines. For this purpose, mixed bilayers of 1-palmitoyl, 2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl, 2-oleoyl-sn-glycero-3-phosphoserine (POPS) were used as model membranes (POPC/POPS 5:1, m/m). Spectra of headgroup- and chain-deuterated POPC and POPS phospholipids, POPC-d4, POPC-d31, POPS-d3, and POPS-d31, were recorded at different temperatures and for various concentrations of the PMP1 fragment. Data obtained from POPS deuterons revealed the formation of specific peptide-POPS complexes giving rise to a slow exchange between free and bound PS lipids, scarcely observed in solid-state NMR studies of lipid-peptide/protein interactions. The stoichiometry of the complex (8 POPS per peptide) was determined and its significance is discussed. The data obtained with headgroup-deuterated POPC were rationalized with a model that integrates the electrostatic perturbation induced by the cationic peptide on the negatively charged membrane interface, and a "spacer" effect due to the intercalation of POPS/PMP1f complexes between choline headgroups.
Roux
M
M
Département de Biologie Cellulaire et Moléculaire, Section de Biophysique des Protéines et des Membranes, CEA and URA CNRS 2096, Centre d'Etudes de Saclay, 91191 Gif sur Yvette Cedex, France. roux@dsvidf.cea.fr
Beswick
V
V
Coïc
Y M
YM
Huynh-Dinh
T
T
Sanson
A
A
Neumann
J M
JM
D000595:Amino Acid Sequence; D055592:Biophysical Phenomena; D001703:Biophysics; D003903:Deuterium; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011510:Proteolipids / Q000737:chemistry* / Q000378:metabolism*; D006180:Proton-Translocating ATPases; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D055672:Static Electricity
D005656:Fungal Proteins; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C489077:PMP1 protein, S cerevisiae; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011510:Proteolipids; D029701:Saccharomyces cerevisiae Proteins; C057561:1-palmitoyl-2-oleoylphosphatidylethanolamine; D003903:Deuterium; D006180:Proton-Translocating ATPases; C028694:1-palmitoyl-2-oleoylphosphatidylcholine