This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Deichaite, I
Right arrow Articles by Resh, M D
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Deichaite, I
Right arrow Articles by Resh, M D

 Previous Article  |  Next Article 

Mol Cell Biol. 1988 October; 8(10): 4295-4301

In vitro synthesis of pp60v-src: myristylation in a cell-free system.

I Deichaite, L P Casson, H P Ling and M D Resh

Department of Biology, Moffett Laboratory, Princeton University, New Jersey 08544.

ABSTRACT

Covalent attachment of myristic acid to pp60v-src, the transforming protein of Rous sarcoma virus, was studied in a cell-free system. Using a synthetic peptide containing the first 11 amino acids of the mature pp60v-src polypeptide sequence as a substrate, we probed lysates from a variety of cells and tissues for N-myristyl transferase (NMT) activity. Nearly every eucaryotic cell type tested contained NMT, including avian, mammalian, insect, and plant cells. Since NMT activity was detected in rabbit reticulocyte lysates, we took advantage of the translational capability of these lysates to determine the precise point during translation at which myristate is attached to pp60v-src. src mRNA, transcribed from cloned v-src DNA, was translated in reticulocyte lysates which had been depleted of endogenous myristate. Addition of [3H]myristate to lysates 10 min after the start of synchronized translation resulted in a dramatic decrease in the incorporation of radiolabeled myristate into pp60v-src polypeptide chains. These results imply that although myristate can be attached posttranslationally to synthetic peptide substrates, myristylation in vivo is apparently a very early cotranslational event which occurs before the first 100 amino acids of the nascent polypeptide chain are polymerized.

Mol Cell Biol. 1988 October; 8(10): 4295-4301




This article has been cited by other articles:

  • Kostiuk, M. A., Corvi, M. M., Keller, B. O., Plummer, G., Prescher, J. A., Hangauer, M. J., Bertozzi, C. R., Rajaiah, G., Falck, J. R., Berthiaume, L. G. (2008). Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue. FASEB J. 22: 721-732 [Abstract] [Full Text]  
  • Raney, A., Kuo, L. S., Baugh, L. L., Foster, J. L., Garcia, J. V. (2005). Reconstitution and Molecular Analysis of an Active Human Immunodeficiency Virus Type 1 Nef/p21-Activated Kinase 2 Complex. J. Virol. 79: 12732-12741 [Abstract] [Full Text]  
  • Ducker, C. E., Upson, J. J., French, K. J., Smith, C. D. (2005). Two N-Myristoyltransferase Isozymes Play Unique Roles in Protein Myristoylation, Proliferation, and Apoptosis. Mol Cancer Res 3: 463-476 [Abstract] [Full Text]  
  • Colombo, S., Longhi, R., Alcaro, S., Ortuso, F., Sprocati, T., Flora, A., Borgese, N. (2005). N-myristoylation determines dual targeting of mammalian NADH-cytochrome b(5) reductase to ER and mitochondrial outer membranes by a mechanism of kinetic partitioning. JCB 168: 735-745 [Abstract] [Full Text]  
  • Haynes, L. P., Thomas, G. M. H., Burgoyne, R. D. (2005). Interaction of Neuronal Calcium Sensor-1 and ADP-ribosylation Factor 1 Allows Bidirectional Control of Phosphatidylinositol 4-Kinase {beta} and trans-Golgi Network-Plasma Membrane Traffic. J. Biol. Chem. 280: 6047-6054 [Abstract] [Full Text]  
  • Korade-Mirnics, Z., Corey, S. J. (2000). Src kinase-mediated signaling in leukocytes. J. Leukoc. Biol. 68: 603-613 [Abstract] [Full Text]  
  • Ishitani, M., Liu, J., Halfter, U., Kim, C.-S., Shi, W., Zhu, J.-K. (2000). SOS3 Function in Plant Salt Tolerance Requires N-Myristoylation and Calcium Binding. Plant Cell 12: 1667-1678 [Abstract] [Full Text]  
  • Qi, Q., Rajala, R. V. S., Anderson, W., Jiang, C., Rozwadowski, K., Selvaraj, G., Sharma, R., Datla, R. (2000). Molecular Cloning, Genomic Organization, and Biochemical Characterization of Myristoyl-CoA:Protein N-Myristoyltransferase from Arabidopsis thaliana. J. Biol. Chem. 275: 9673-9683 [Abstract] [Full Text]  
  • Hof, W., Resh, M. D. (1999). Dual Fatty Acylation of p59Fyn Is Required for Association with the T Cell Receptor {zeta} Chain through Phosphotyrosine-Src Homology Domain-2 Interactions. JCB 145: 377-389 [Abstract] [Full Text]  
  • Hermida-Matsumoto, L., Resh, M. D. (1999). Human Immunodeficiency Virus Type 1 Protease Triggers a Myristoyl Switch That Modulates Membrane Binding of Pr55gag and p17MA. J. Virol. 73: 1902-1908 [Abstract] [Full Text]  
  • Sarkar, S. N., Bandyopadhyay, S., Ghosh, A., Sen, G. C. (1999). Enzymatic Characteristics of Recombinant Medium Isozyme of 2'-5' Oligoadenylate Synthetase. J. Biol. Chem. 274: 1848-1855 [Abstract] [Full Text]  
  • Giang, D. K., Cravatt, B. F. (1998). A Second Mammalian N-Myristoyltransferase. J. Biol. Chem. 273: 6595-6598 [Abstract] [Full Text]  
  • Glover, C. J., Hartman, K. D., Felsted, R. L. (1997). Human N-Myristoyltransferase Amino-terminal Domain Involved in Targeting the Enzyme to the Ribosomal Subcellular Fraction. J. Biol. Chem. 272: 28680-28689 [Abstract] [Full Text]  
  • Hof, W. v. t, Resh, M. D. (1997). Rapid Plasma Membrane Anchoring of Newly Synthesized p59 fyn: Selective Requirement for NH2-Terminal Myristoylation and Palmitoylation at Cysteine-3. JCB 136: 1023-1035 [Abstract] [Full Text]  
  • Ntwasa, M, Egerton, M, Gay, N. (1997). Sequence and expression of Drosophila myristoyl-CoA: protein N-myristoyl transferase: evidence for proteolytic processing and membrane localisation. J. Cell Sci. 110: 149-156 [Abstract]  
  • Saouaf, S. J., Kut, S. A., Fargnoli, J., Rowley, R. B., Bolen, J. B., Mahajan, S. (1995). Reconstitution of the B Cell Antigen Receptor Signaling Components in COS Cells. J. Biol. Chem. 270: 27072-27078 [Abstract] [Full Text]  
  • David-Pfeuty, T, Bagrodia, S, Shalloway, D (1993). Differential localization patterns of myristoylated and nonmyristoylated c-Src proteins in interphase and mitotic c-Src overexpresser cells. J. Cell Sci. 105: 613-628 [Abstract]  
  • Utsumi, T., Sato, M., Nakano, K., Takemura, D., Iwata, H., Ishisaka, R. (2001). Amino Acid Residue Penultimate to the Amino-terminal Gly Residue Strongly Affects Two Cotranslational Protein Modifications, N-Myristoylation and N-Acetylation. J. Biol. Chem. 276: 10505-10513 [Abstract] [Full Text]  
  • Austin, C., Hinners, I., Tooze, S. A. (2000). Direct and GTP-dependent Interaction of ADP-ribosylation Factor 1 with Clathrin Adaptor Protein AP-1 on Immature Secretory Granules. J. Biol. Chem. 275: 21862-21869 [Abstract] [Full Text]  
  • Hunter, C., Sung, P., Schejter, E. D., Wieschaus, E. (2002). Conserved Domains of the Nullo Protein Required for Cell-Surface Localization and Formation of Adherens Junctions. Mol. Biol. Cell 13: 146-157 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»