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

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dc.contributor.authorNavarro-Lerida, Inmaculada
dc.contributor.authorAlvarez-Barrientos, Alberto
dc.contributor.authorGavilanes, Francisco
dc.contributor.authorRodriguez-Crespo, Ignacio
dc.contributor.funderComunidad de Madrid (España)
dc.contributor.funderSpanish DGI
dc.date.accessioned2021-02-17T07:06:29Z
dc.date.available2021-02-17T07:06:29Z
dc.date.issued2002-08-01
dc.description.abstractUsing 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.es_ES
dc.description.peerreviewedSíes_ES
dc.format.numberPt 15es_ES
dc.format.page3119-30es_ES
dc.format.volume115es_ES
dc.identifier.citationJ Cell Sci. 2002; 115(Pt 15):3119-30es_ES
dc.identifier.doi10.1242/jcs.209973es_ES
dc.identifier.issn0021-9533
dc.identifier.journalJournal of cell sciencees_ES
dc.identifier.pubmedID12118067es_ES
dc.identifier.urihttp://hdl.handle.net/20.500.12105/11915
dc.language.isoenges_ES
dc.publisherThe Company of Biologistses_ES
dc.relation.projectIDinfo:eu-repo/grantAgreement/ES/08.4/0039.1/2000es_ES
dc.relation.projectIDinfo:eu-repo/grantAgreement/ES/2000-0545es_ES
dc.relation.publisherversionhttps://doi.org/10.1242/jcs.209973es_ES
dc.repisalud.institucionCNICes_ES
dc.repisalud.orgCNICCNIC::Grupos de investigación::Antiguos CNICes_ES
dc.rights.accessRightsopen accesses_ES
dc.subject.meshRecombinant Fusion Proteinses_ES
dc.subject.meshAmino Acid Sequencees_ES
dc.subject.meshAnimalses_ES
dc.subject.meshCOS Cellses_ES
dc.subject.meshCaveolin 1es_ES
dc.subject.meshCaveolinses_ES
dc.subject.meshCell Compartmentationes_ES
dc.subject.meshCell Membranees_ES
dc.subject.meshCholesteroles_ES
dc.subject.meshCytoplasmes_ES
dc.subject.meshEukaryotic Cellses_ES
dc.subject.meshGolgi Apparatuses_ES
dc.subject.meshGreen Fluorescent Proteinses_ES
dc.subject.meshLuminescent Proteinses_ES
dc.subject.meshMembrane Microdomainses_ES
dc.subject.meshMicroscopy, Confocales_ES
dc.subject.meshMyristic Acides_ES
dc.subject.meshOctoxynoles_ES
dc.subject.meshPalmitateses_ES
dc.subject.meshPalmitic Acides_ES
dc.subject.meshProtein Transportes_ES
dc.subject.meshProteinses_ES
dc.subject.meshSolubilityes_ES
dc.subject.meshSphingomyelinses_ES
dc.titleDistance-dependent cellular palmitoylation of de-novo-designed sequences and their translocation to plasma membrane subdomains.es_ES
dc.typejournal articlees_ES
dc.type.hasVersionVoRes_ES
dspace.entity.typePublication
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