Acylation of Proteins with Myristic Acid Occurs Cotranslationally

Wilcox, Celeste A.; Hu, Jing; Olson, Eric N.

doi:10.1126/science.3685978

Cited by 264 publications

(164 citation statements)

References 22 publications

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“…Since both NMTs and methionine aminopeptidases function cotranslationally while the nascent polypeptide chain is still attached to the ribosomes (20,36,37), it might be possible to speculate that these two enzymes may share similar restrictions with respect to their substrate specificities.…”

Section: Discussionmentioning

confidence: 99%

Amino Acid Residue Penultimate to the Amino-terminal Gly Residue Strongly Affects Two Cotranslational Protein Modifications, N-Myristoylation andN-Acetylation

Utsumi

Sato

Nakano

et al. 2001

Journal of Biological Chemistry

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To examine the amino-terminal sequence requirements for cotranslational protein N-myristoylation, a series of site-directed mutagenesis of N-terminal region were performed using tumor necrosis factor as a nonmyristoylated model protein. Subsequently, the susceptibility of these mutants to protein N-myristoylation was evaluated by metabolic labeling in an in vitro translation system or in transfected cells. It was found that the amino acid residue at position 3 in an N-myristoylation consensus motif, Met-Gly-X-X-X-Ser-X-X-X, strongly affected the susceptibility of the protein to two different cotranslational protein modifications, N-myristoylation and N-acetylation; 10 amino acids (Ala, Ser, Cys, Thr, Val, Asn, Leu, Ile, Gln, and His) with a radius of gyration smaller than 1.80 Å directed N-myristoylation, two negatively charged residues (Asp and Glu) directed N-acetylation, and two amino acids (Gly and Met) directed heterogeneous modification with both N-myristoylation and N-acetylation. The amino acid requirements at this position for the two modifications were dramatically changed when Ser at position 6 in the consensus motif was replaced with Ala. Thus, the amino acid residue penultimate to the N-terminal Gly residue strongly affected two cotranslational protein modifications, N-myristoylation and N-acetylation, and the amino acid requirements at this position for these two modifications were significantly affected by downstream residues.

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Section: Discussionmentioning

confidence: 99%

Amino Acid Residue Penultimate to the Amino-terminal Gly Residue Strongly Affects Two Cotranslational Protein Modifications, N-Myristoylation andN-Acetylation

Utsumi

Sato

Nakano

et al. 2001

Journal of Biological Chemistry

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“…This characteristic differentiates palmitoylation from myristoylation, a cotranslational modification that is blocked by protein synthesis inhibitors (27). To investigate the cellular mechanism of caveolin-1 palmitoylation, bovine aortic EC were pretreated with cycloheximide for 60 min before incubation with (28).…”

Section: Incorporation Of Palmitate Into Caveolin-1 Requires De Novomentioning

confidence: 99%

Palmitoylation of Caveolin-1 in Endothelial Cells Is Post-translational but Irreversible

Parat

Fox

2001

Journal of Biological Chemistry

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Caveolin-1 is a palmitoylated protein involved in assembly of signaling molecules in plasma membrane subdomains termed caveolae and in intracellular cholesterol transport. Three cysteine residues in the C terminus of caveolin-1 are subject to palmitoylation, which is not necessary for caveolar targeting of caveolin-1. Protein palmitoylation is a post-translational and reversible modification that may be regulated and that in turn may regulate conformation, membrane association, protein-protein interactions, and intracellular localization of the target protein. We have undertaken a detailed analysis of [ 3 H]palmitate incorporation into caveolin-1 in aortic endothelial cells. The linkage of palmitate to caveolin-1 was hydroxylamine-sensitive and thus presumably a thioester bond. However, contrary to expectations, palmitate incorporation was blocked completely by the protein synthesis inhibitors cycloheximide and puromycin. In parallel experiments to show specificity, palmitoylation of aortic endothelial cell-specific nitric-oxide synthase was unaffected by these reagents. Inhibitors of protein trafficking, brefeldin A and monensin, blocked caveolin-1 palmitoylation, indicating that the modification was not cotranslational but rather required caveolin-1 transport from the endoplasmic reticulum and Golgi to the plasma membrane. In addition, immunophilin chaperones that form complexes with caveolin-1, i.e. FK506-binding protein 52, cyclophilin A, and cyclophilin 40, were not necessary for caveolin-1 palmitoylation because agents that bind immunophilins did not inhibit palmitoylation. Pulse-chase experiments showed that caveolin-1 palmitoylation is essentially irreversible because the release of [ Originally named to describe membrane invaginations at the cell surface, caveolae are specialized plasmalemmal domains rich in glycosphingolipids, cholesterol, and lipid-anchored membrane proteins. In endothelial cells (EC) 1 , caveolae have a striated coat, the major protein of which is caveolin-1 (1). Caveolin-1 is involved in cholesterol trafficking (2), oligomerizes to form a scaffold for assembly of signaling molecules including receptors, signal transducers, and effectors (3), and regulates the activation state of these signaling complexes (4).Lipid modification, including palmitoylation, is an important mechanism targeting signaling proteins to caveolae. Among the palmitoylated proteins in caveolae are heterotrimeric Gprotein ␣-subunits (e.g. ␣ s , ␣ i , ␣ o , and ␣ q ), G-protein linked receptors, regulator of G-protein signaling proteins, p21 ras , nonreceptor tyrosine kinases, and EC-specific nitric-oxide synthase (eNOS) (5). Palmitoylation of several of these proteins has been shown to be a post-translational and reversible modification that is also subject to regulation (6). The post-translational nature of palmitoylation has been shown by cell culture experiments in which palmitate labeling occurs even in the presence of protein synthesis inhibitors (7). These experiments clearly distinguish protein palmitoyl...

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“…Lipid membrane domains (also known as lipid rafts) are organized domains of the plasma membrane and other intracellular membranes and can be viewed as signaling platforms that serve to colocalize molecular components, facilitating their interaction and supporting signaling (25)(26)(27)(28). Several raft-associated proteins are anchored to the cytosolic surface of the raft plasma membrane by acylation with myristate and palmitate and isoprenylation with farnesyl or geranylgeranyl moieties (29)(30)(31)(32)(33). Because myristic and palmitic acids induce dome formation in LA7 cells (15), we investigated the possible association of rat8 to lipid membrane domains.…”

mentioning

confidence: 99%

Association of rat8 with Fyn protein kinase via lipid rafts is required for rat mammary cell differentiationin vitro

Zucchi

Prinetti

Scotti

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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We previously identified rat8 in the pathway involved in epithelial cell differentiation that occurs in the rat mammary gland at pregnancy when tubules and alveoli are formed. rat8, which encodes an IFN-inducible membrane protein, is the rat homologue of the mouse gene fragilis. By differential detergent extraction and isopycnic sucrose density gradients, we show that rat8 protein is associated to lipid membrane domains together with Lyn and Fyn, members of the Src tyrosine kinase family. We also show that recruitment of rat8 to lipid membrane domains is a necessary step in mammary epithelial cell differentiation. Immunoprecipitation analysis, performed with an anti-Fyn protein antibody, shows that rat8 was present in the Fyn immunoprecipitate. Antisense oligonucleotides, used to inhibit Fyn protein expression, block mammary cell differentiation. Taken together, these results suggest that the functional interaction, via lipid membrane domains, of rat8 and Fyn proteins is required for mammary cell differentiation. Therefore, rat8, like fragilis, may be involved in developmental decisions and the demarcation of a subset of cells in the mammary gland that cause epithelial cells to develop into a network of tubuloalveolar structures involved in secretion.

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Acylation of Proteins with Myristic Acid Occurs Cotranslationally

Cited by 264 publications

References 22 publications

Amino Acid Residue Penultimate to the Amino-terminal Gly Residue Strongly Affects Two Cotranslational Protein Modifications, N-Myristoylation andN-Acetylation

Amino Acid Residue Penultimate to the Amino-terminal Gly Residue Strongly Affects Two Cotranslational Protein Modifications, N-Myristoylation andN-Acetylation

Palmitoylation of Caveolin-1 in Endothelial Cells Is Post-translational but Irreversible

Association of rat8 with Fyn protein kinase via lipid rafts is required for rat mammary cell differentiationin vitro

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