QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

This Article Figures Only Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Navarro-Lérida, I. Articles by Rodriguez-Crespo, I. Search for Related Content PubMed PubMed Citation Articles by Navarro-Lérida, I. Articles by Rodriguez-Crespo, I.

Distance-dependent cellular palmitoylation of de-novo-designed sequences and their translocation to plasma membrane subdomains

¹ Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas, Universidad Complutense, 28040 Madrid, Spain
² Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain

^* Author for correspondence (e-mail: nacho{at}bbm1.ucm.es )

Accepted 17 May 2002

Using recursive PCR, we created an artificial protein sequence that consists of a consensus myristoylation motif (MGCTLS) followed by the triplet AGS repeated nine times and fused to the GFP reporter. This linker-GFP sequence was utilized as a base to produce multiple mutants that were used to transfect COS-7 cells. Constructs where a `palmitoylable' cysteine residue was progressively moved apart from the myristoylation site to positions 3, 9, 15 and 21 of the protein sequence were made, and these mutants were used to investigate the effect of protein myristoylation on subsequent palmitoylation, subcellular localization, membrane association and caveolin-1 colocalization. In all cases, dual acylation of the GFP chimeras correlated with translocation to Triton X-100-insoluble cholesterol/sphingomyelin-enriched subdomains. Whereas a strong Golgi labeling was observed in all the myristoylated chimeras, association with the plasma membrane was only observed in the dually acylated constructs. Taking into account the conflicting data regarding the existence and specificity of cellular palmitoyl-transferases, our results provide evidence that de-novo-designed sequences can be efficiently S-acylated with palmitic acid in vivo, strongly supporting the hypothesis that non-enzymatic protein palmitoylation can occur within mammalian cells. Additionally, this palmitoylation results in the translocation of the recombinant construct to low-fluidity domains in a myristate-palmitate distance-dependent manner.

Key words: Myristoylation, Caveolin, Palmitoylation, Rafts, Subcellular targeting

This article has been cited by other articles:

				S. C. Lee, W.-Z. Lan, B.-G. Kim, L. Li, Y. H. Cheong, G. K. Pandey, G. Lu, B. B. Buchanan, and S. Luan A protein phosphorylation/dephosphorylation network regulates a plant potassium channel PNAS, October 2, 2007; 104(40): 15959 - 15964. [Abstract] [Full Text] [PDF]

				I. Navarro-Lerida, M. Martinez-Moreno, I. Ventoso, A. Alvarez-Barrientos, and I. Rodriguez-Crespo Binding of CAP70 to Inducible Nitric Oxide Synthase and Implications for the Vectorial Release of Nitric Oxide in Polarized Cells Mol. Biol. Cell, July 1, 2007; 18(7): 2768 - 2777. [Abstract] [Full Text] [PDF]

				K. Subramanian, L. E. P. Dietrich, H. Hou, T. J. LaGrassa, C. T. A. Meiringer, and C. Ungermann Palmitoylation determines the function of Vac8 at the yeast vacuole J. Cell Sci., June 15, 2006; 119(12): 2477 - 2485. [Abstract] [Full Text] [PDF]

				I. Navarro-Lerida, A. Alvarez-Barrientos, and I. Rodriguez-Crespo N-terminal palmitoylation within the appropriate amino acid environment conveys on NOS2 the ability to progress along the intracellular sorting pathways J. Cell Sci., April 15, 2006; 119(8): 1558 - 1569. [Abstract] [Full Text] [PDF]

				O. Rocks, A. Peyker, M. Kahms, P. J. Verveer, C. Koerner, M. Lumbierres, J. Kuhlmann, H. Waldmann, A. Wittinghofer, and P. I. H. Bastiaens An Acylation Cycle Regulates Localization and Activity of Palmitoylated Ras Isoforms Science, March 18, 2005; 307(5716): 1746 - 1752. [Abstract] [Full Text] [PDF]

				I. Navarro-Lerida, M. M. Corvi, A. A. Barrientos, F. Gavilanes, L. G. Berthiaume, and I. Rodriguez-Crespo Palmitoylation of Inducible Nitric-oxide Synthase at Cys-3 Is Required for Proper Intracellular Traffic and Nitric Oxide Synthesis J. Biol. Chem., December 31, 2004; 279(53): 55682 - 55689. [Abstract] [Full Text] [PDF]

				X. Ren, A. G. Ostermeyer, L. T. Ramcharan, Y. Zeng, D. M. Lublin, and D. A. Brown Conformational Defects Slow Golgi Exit, Block Oligomerization, and Reduce Raft Affinity of Caveolin-1 Mutant Proteins Mol. Biol. Cell, October 1, 2004; 15(10): 4556 - 4567. [Abstract] [Full Text] [PDF]

				I. Navarro-Lerida, M. T. Portoles, A. A. Barrientos, F. Gavilanes, L. Bosca, and I. Rodriguez-Crespo Induction of nitric oxide synthase-2 proceeds with the concomitant downregulation of the endogenous caveolin levels J. Cell Sci., May 1, 2004; 117(9): 1687 - 1697. [Abstract] [Full Text] [PDF]