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Fatty Acylation of Proteins: New Insights Into Membrane Targeting of Myristoylated and Palmitoylated Proteins

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Fatty Acylation of Proteins: New Insights Into Membrane Targeting of Myristoylated and Palmitoylated Proteins View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biochimica et Biophysica Acta 1451 (1999) 1^16 www.elsevier.com/locate/bba Review Fatty acylation of proteins: new insights into membrane targeting of myristoylated and palmitoylated proteins Marilyn D. Resh * Cell Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 143, New York, NY 10021, USA Received 15 March 1999; received in revised form 20 May 1999; accepted 2 June 1999 Abstract Covalent attachment of myristate and/or palmitate occurs on a wide variety of viral and cellular proteins. This review will highlight the latest advances in our understanding of the enzymology of N-myristoylation and palmitoylation as well as the functional consequences of fatty acylation of key signaling proteins. The role of myristate and palmitate in promoting membrane binding as well as specific membrane targeting will be reviewed, with emphasis on the Src family of tyrosine protein kinases and K subunits of heterotrimeric G proteins. The use of myristoyl switches and regulated depalmitoylation as mechanisms for achieving reversible membrane binding and regulated signaling will also be explored. ß 1999 Elsevier Science B.V. All rights reserved. Keywords: Fatty acylation; Myristoylation; Palmitoylation; Membrane proteins; Membrane rafts Contents 1. Introduction .......................................................... 2 2. Protein N-myristoylation ................................................. 2 2.1. Structure and function of N-myristoyl transferase ........................... 2 2.2. Functions of protein N-myristoylation . .................................. 4 2.3. Regulation of membrane binding of N-myristoylated proteins .................. 6 3. Protein palmitoylation . ................................................. 8 3.1. Enzymology of palmitoylation . ....................................... 8 3.2. The role of palmitoylation in membrane binding and plasma membrane targeting . 11 4. Conclusions and perspectives . ............................................ 13 References ............................................................... 14 * Fax: +1-212-717-3317; E-mail: [email protected] 0167-4889 / 99 / $ ^ see front matter ß 1999 Elsevier Science B.V. All rights reserved. PII: S0167-4889(99)00075-0 BBAMCR 14503 30-7-99 2 M.D. Resh / Biochimica et Biophysica Acta 1451 (1999) 1^16 1. Introduction tein substrates. To date, nearly one dozen NMTs from fungal and mammalian sources have been iden- Covalent attachment of fatty acids to proteins is ti¢ed. Studies of NMT from the yeast S. cerevisiae now a widely recognized form of protein modi¢- reveal that the catalytic cycle exhibits the following cation. Many fatty acylated proteins play key roles Bi Bi reaction mechanism [6]: (1) myristoyl CoA in regulating cellular structure and function. Within binds to NMT; (2) the peptide substrate binds to the past 3 years, several reviews have focused on NMT; (3) myristate is transferred to the N-terminal fatty acylation of signaling proteins [1^4]. This re- glycine of the peptide; (4) CoA is released from the view will highlight some of the newest advances in enzyme; (5) myristoyl-peptide is released. In cells, N- protein fatty acylation, particularly with regard to myristoylation is a cotranslational process that oc- membrane binding and targeting of fatty acylated curs while the nascent polypeptide chain is still at- proteins. tached to the ribosome [7,8]. The two most common forms of protein fatty acyl- The requirements for both fatty acid and peptide ation are modi¢cation with myristate, a 14-carbon substrate speci¢cities of NMT have been character- saturated fatty acid, and palmitate, a 16-carbon sa- ized and have been extensively reviewed [9]. Myris- turated fatty acid. The enzymology of the myristoyl- tate is attached speci¢cally to the N-terminus of the ation reaction is well understood. Proteins that are acceptor protein via an amide bond. Longer or destined to become myristoylated begin with the se- shorter fatty acyl CoAs are poor substrates, and their quence: Met^Gly. The initiating methionine is re- corresponding fatty acids are generally not trans- moved cotranslationally by methionine amino-pepti- ferred by NMT to acceptor proteins. It is interesting dase, and myristate is linked to Gly-2 via an amide to note that palmitoyl CoA binds to NMT with bond. As discussed below, N-myristoylation is cata- approximately the same Km as myristoyl CoA, lyzed by N-myristoyl transferase, an enzyme that has although palmitate is not transferred by NMT to recently been extensively characterized. In contrast, proteins. Since the concentration of palmitoyl CoA the enzymology of palmitoylation reaction(s) is more is 5^20-fold higher than myristoyl CoA in most complex and is poorly understood. Nearly all palmi- cells, a mechanism must exist to prevent palmitoyl toylated proteins are acylated by attachment of pal- CoA from functioning as a competitive inhibitor of mitate through a thioester linkage to the sulfhydryl NMT in vivo. It has been proposed that due to its group of cysteine. (One exception is Sonic hedgehog, higher hydrophobicity, most of the palmitoyl CoA is a protein that contains palmitate linked to the amino sequestered in cell membranes and therefore is not group of the N-terminal cysteine [5].) The location of accessible to cytoplasmic NMT [9,10]. these palmitoylated cysteine residues varies-some are Several N-myristoylated proteins, including recov- present near the N- or C-termini of proteins, while erin, transducin and protein kinase A, have been others are located near transmembrane domains. shown to be heterogeneously fatty acylated, contain- Both myristoylation and palmitoylation can be dy- ing 12:0, 14:1 and 14:2 fatty acids in addition to namically regulated: the myristate moiety can be se- 14:0 myristate [11,12]. Examination of N-myristoyl- questered through the use of myristoyl switches, ated proteins in di¡erent tissues reveals that hetero- while palmitate can be removed by protein palmitoyl genous N-acylation is primarily restricted to the ret- thioesterases. ina [9,13]. The reason for the retina-speci¢c nature of heterogenous acylation is not clear, as the fatty acyl CoA pools in retina appear nearly identical to those 2. Protein N-myristoylation in other tissues [13]. The consensus sequence for NMT protein sub- 2.1. Structure and function of N-myristoyl transferase strates is: Met^Gly^X^X^X^Ser/Thr^. The initiating Met is removed by methionine amino peptidase dur- N-myristoyl transferase (NMT) is a 50^60 kDa ing translation and Gly-2 becomes the N-terminal monomeric enzyme that catalyzes transfer of myris- amino acid. The requirement for Gly at the N-termi- tate from myristoyl-CoA to suitable peptide and pro- nus is absolute; no other amino acid will substitute. BBAMCR 14503 30-7-99 M.D. Resh / Biochimica et Biophysica Acta 1451 (1999) 1^16 3 Table 1 Table 1 (continued) N-Myristoyolated proteins: sequence and function 4. Membrane- and cytoskeletal-bound structural proteins 1. Protein kinases and phosphatases MARCKS M G AQFS KTAAKGEATA Src family tyrosine kinases MAC-MARCKS M G SQSS KAPRGDVTAE Src M G SSKS KPKDPSQRRR Annexin XIII M G NRHAKASSPQGFDV Yes M G CIKS KEDKGPAMKY Rapsyn M G QDQT KQQIEKGLQL Fyn M G CVQCKDKEATKLTE Pallidin (Band 4.2) M G QALS IKSCDFHAAE Lyn M G CIKS KRKDNLNDDE Hisactophilin 2 M G NRAFKAHNGHYLSA Lck M G CVCS SNPEDDWMEN Hck M G CMKS KFLQVGGNTG 5. Viral proteins Fgr M G CVFCKKLEPVATAK Gag proteins Yrk M G CVHCKEKISGKGQG HIV-1 M G ARAS VLSGGELDRW Blk M G LLSS KRQVSEKGKG Moloney murine M G QTVT TPLSLTLDHW Abl tyrosine kinases sarcoma virus c-Abl M G QQPGKVLGDQRRPS Friend murine M G QAVT TPLSLTLDHW Arg M G QQVGRVGEAPGLQQ leukemia virus Serine/threonine kinases and anchoring proteins Bovine leukemia virus M G NSPS YNPPAGISPS cAMP-Dependent Baboon endogenous M G QTLT TPLSLTLTHF protein kinase, virus catalytic subunit Feline sarcoma virus M G QTIT TPLSLTLDHW Alpha M G NAAAAKKGSEQESV (G-R) Beta-1 M G NAAT AKKGSEVESV FBR murine M G QTVT TPLSLTLEHW Yeast protein kinase M G AQLS LVVQASPSIA osteosarcoma virus VPS15 Mason^P¢zer monkey M G QELS QHERYVEQLK MPSK M G HALCVCSRGTVIID virus AKAP18 M G QLCCFPFSRDEGKI Mouse mammary M G VSGS KGQKLFVSVL Phosphatases tumor virus Calcineurin B M G NEAS YPLEMCSHFD Other viral proteins Yeast phosphatase M G NSGS KQHTKHNSKK HIV-1 Nef M G GKWS KSSVVGWPTV PP-Z2 Duck hepatitis B virus M G QHPAKSMDVRRIEG Rhinovirus 16 M G AQVS RQNVGTHSTQ 2. Guanine nucleotide binding proteins Herpes simplex virus M G LAFS GARPCCCRHN GK proteins Ul11 GKi1 M G CTLS AEDKAAVERS African swine fever M G NRGS STSSRPPLSS GKo M G CTLS AEERAALERS virus pp220 GKt M G AGAS AEEKHSRELE Chlorella virus Vp260 M G SYFVPPANYFFKDI GKx M G CRQS SEEKEAARRS ADP-ribosylation factors 6. Miscellaneous Arf-1 M G LSFT KLFSRLFAKK NADH cytochrome b5 M G AQLS TLSRVVLSPV Arf-3 M G NIFGNLLKSLIGKK reductase Arf-5 M G LTVS ALFSRIFGKK Nitric oxide synthase M G NLKS VGQEPGPPCG Arf-6 M G KVLS KIFGNKEMRI Enteropeptidase M G SKRS VPSRHRSLTT precursor 3. Ca2 binding/EF hand proteins (2P,5P) OligoA M G NGES QLSSVPAQKL Recoverin M G NSKS GALSKEILEE synthetase Neurocalcin M G KQNS KLRPEVMQDL NADH-Ubiquinone M G AHLVRRYLGDASVE Aplycalcin M G KRAS KLKPEEVEEL Oxidoreductase Visinin-like protein 3 M G KQNS KLRPEVLQDL BASP1 M G GKLS KKKKGYNVND Ca2 binding protein M G SRAS TLLRDEELEE FRS2 M G SCCS CPDKDTVPDN P22 cPLA2-Q M G SSEVSIIPGLQKEE NAP-22 M G SKLS KKKKGYNVND Hippocalcin M G KQNS KLRPEMLQDL GCAP 1 M G NVMEGKSVEELSST GCAP 2 M G QQFS WEEAEENGAV S-Modulin M G NTKS GALSKEILEE Rem-1 M G KQNS KLRPEVLQDL BBAMCR 14503 30-7-99 4 M.D. Resh / Biochimica et Biophysica Acta 1451 (1999) 1^16 However, not all proteins with an N-terminal glycine binds in a conformation similar to a question mark are N-myristoylated and the ability to be recognized and, together with C-terminal regions of NMT, by NMT depends on the downstream amino acid forms a portion of the peptide binding site.
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