title: S-palmitoylation and s-oleoylation of rabbit and pig sarcolipin.
abstract: Sarcolipin (SLN) is a regulatory peptide present in sarcoplasmic reticulum (SR) from skeletal muscle of animals. We find that native rabbit SLN is modified by a fatty acid anchor on Cys-9 with a palmitic acid in about 60% and, surprisingly, an oleic acid in the remaining 40%. SLN used for co-crystallization with SERCA1a (Winther, A. M., Bublitz, M., Karlsen, J. L., Moller, J. V., Hansen, J. B., Nissen, P., and Buch-Pedersen, M. J. (2013) Nature 495, 265-2691; Ref. 1) is also palmitoylated/oleoylated, but is not visible in crystal structures, probably due to disorder. Treatment with 1 m hydroxylamine for 1 h removes the fatty acids from a majority of the SLN pool. This treatment did not modify the SERCA1a affinity for Ca(2+) but increased the Ca(2+)-dependent ATPase activity of SR membranes indicating that the S-acylation of SLN or of other proteins is required for this effect on SERCA1a. Pig SLN is also fully palmitoylated/oleoylated on its Cys-9 residue, but in a reverse ratio of about 40/60. An alignment of 67 SLN sequences from the protein databases shows that 19 of them contain a cysteine and the rest a phenylalanine at position 9. Based on a cladogram, we postulate that the mutation from phenylalanine to cysteine in some species is the result of an evolutionary convergence. We suggest that, besides phosphorylation, S-acylation/deacylation also regulates SLN activity.
authors.affiliation: From the Laboratoire des Protéines Membranaires, UMR 8221, Commissariat à l'Energie Atomique (CEA), Université Paris-Sud and Centre National de la Recherche Scientifique (CNRS), F91191, Gif-sur-Yvette, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Montigny
authors.affiliation: Institut de Biochimie et Biophysique Moléculaire et Cellulaire, CNRS UMR 8619, Université Paris-Sud, F91400, Orsay, France.
authors.forename: Paulette
authors.initials: P
authors.lastname: Decottignies
authors.affiliation: Institut de Biochimie et Biophysique Moléculaire et Cellulaire, CNRS UMR 8619, Université Paris-Sud, F91400, Orsay, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Le Maréchal
authors.affiliation: Laboratoire Evolution, Génomes et Spéciation, CNRS UPR 9034, Centre de Recherche de Gif and Université Paris-Sud, F91190, Gif-sur-Yvette, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Capy
authors.affiliation: Centre for Membrane Pumps in Cells and Disease, PUMPKIN, Danish National Research Foundation, Department of Molecular Biology and Genetics, and.
authors.forename: Maike
authors.initials: M
authors.lastname: Bublitz
authors.affiliation: Centre for Membrane Pumps in Cells and Disease, PUMPKIN, Danish National Research Foundation, Department of Molecular Biology and Genetics, and Department of Biomedicine, Aarhus University, 8000, Aarhus, Denmark.
authors.forename: Claus
authors.initials: C
authors.lastname: Olesen
authors.affiliation: Centre for Membrane Pumps in Cells and Disease, PUMPKIN, Danish National Research Foundation, Department of Molecular Biology and Genetics, and.
authors.forename: Jesper Vuust
authors.initials: JV
authors.lastname: Møller
authors.affiliation: Centre for Membrane Pumps in Cells and Disease, PUMPKIN, Danish National Research Foundation, Department of Molecular Biology and Genetics, and.
authors.forename: Poul
authors.initials: P
authors.lastname: Nissen
authors.affiliation: From the Laboratoire des Protéines Membranaires, UMR 8221, Commissariat à l'Energie Atomique (CEA), Université Paris-Sud and Centre National de la Recherche Scientifique (CNRS), F91191, Gif-sur-Yvette, France, marc.lemaire@cea.fr.
authors.forename: Marc
authors.initials: M
authors.lastname: le Maire
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D005075:Biological Evolution; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D019811:Hydroxylamine / Q000737:chemistry; D007700:Kinetics; D054878:Lipoylation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism; D018482:Muscle, Skeletal / Q000737:chemistry / Q000378:metabolism*; D019301:Oleic Acid / Q000737:chemistry* / Q000378:metabolism; D019308:Palmitic Acid / Q000737:chemistry* / Q000378:metabolism; D010649:Phenylalanine / Q000737:chemistry* / Q000378:metabolism; D010802:Phylogeny; D011499:Protein Processing, Post-Translational; D011510:Proteolipids / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D012519:Sarcoplasmic Reticulum; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013045:Species Specificity; D013552:Swine; D013816:Thermodynamics
chemical_list: D009124:Muscle Proteins; D011510:Proteolipids; C077258:sarcolipin; D019811:Hydroxylamine; D019301:Oleic Acid; D019308:Palmitic Acid; D010649:Phenylalanine; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D003545:Cysteine
keywords: Calcium ATPase; Mass Spectrometry (MS); Membrane Protein; Protein Acylation; Protein Oleoylation; Protein Palmitoylation; Sarcolipin; Sarcoplasmic Reticulum (SR)