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J Biol Chem. 2014 Dec 5;289(49):33850-61. doi: 10.1074/jbc.M114.590307. Epub 2014 Oct 9.

S-palmitoylation and s-oleoylation of rabbit and pig sarcolipin.

Author information

1
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.
2
Institut de Biochimie et Biophysique Moléculaire et Cellulaire, CNRS UMR 8619, Université Paris-Sud, F91400, Orsay, France.
3
Laboratoire Evolution, Génomes et Spéciation, CNRS UPR 9034, Centre de Recherche de Gif and Université Paris-Sud, F91190, Gif-sur-Yvette, France.
4
Centre for Membrane Pumps in Cells and Disease, PUMPKIN, Danish National Research Foundation, Department of Molecular Biology and Genetics, and.
5
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.
6
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.

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.

KEYWORDS:

Calcium ATPase; Mass Spectrometry (MS); Membrane Protein; Protein Acylation; Protein Oleoylation; Protein Palmitoylation; Sarcolipin; Sarcoplasmic Reticulum (SR)

PMID:
25301946
PMCID:
PMC4256321
DOI:
10.1074/jbc.M114.590307
[Indexed for MEDLINE]
Free PMC Article

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