1. Antibodies to an NH2-terminal myristoyl glycine moiety can detect NH2-terminal myristoylated proteins in the retrovirus-infected cells
S Shoji, A Tashiro, K Furuishi, O Takenaka, Y Kida, S Horiuchi, T Funakoshi, Y Kubota Biochem Biophys Res Commun. 1989 Jul 31;162(2):724-32. doi: 10.1016/0006-291x(89)92370-x.
Novel antibodies were raised against a synthetic NH2-terminal myristoyl glycine moiety which is characteristic of N-myristoyl-proteins. Antisera raised against N-myristoyl-Gly-hemocyanin reacted with N-myristoyl-Gly-[125I]albumin. The immunoreaction was competed for by albumin conjugated with N-myristoyl-glycine, while underivatized albumin had no effect. Of the [3H]myristate-labeled proteins detected, pp60v-src, which is a transforming protein of Rous sarcoma virus, and p19gag and p17gag, which are core proteins in the human T-cell leukemia virus and the human immunodeficiency virus, were identified as N-myristoylated proteins by the radioimmunoprecipitation analyses with the antibody.
2. The HIV-1 Gag precursor Pr55gag synthesized in yeast is myristoylated and targeted to the plasma membrane
E Jacobs, D Gheysen, D Thines, M Francotte, M de Wilde Gene. 1989 Jun 30;79(1):71-81. doi: 10.1016/0378-1119(89)90093-0.
Myristoylation of the Pr65gag protein from Moloney murine leukemia virus has been shown to be essential for virus particle formation [Rein et al., Proc. Natl. Acad. Sci. USA 83 (1986) 7246-7250], and by analogy, myristoylation of the human immunodeficiency virus (HIV) Gag precursor could possibly play a similar role. We have investigated the expression and myristoylation of the complete HIV Gag precursor Pr55gag in yeast, the subcellular localization of that protein, and the contribution of the myristoyl-glycine residue to this localization. Immunogold labelling of myristoylated Pr55gage with antibodies directed against HIV Gag products was apparent in the vicinity of the plasma membrane. On the contrary, non-myristoylated derivatives of Pr55gag were only detected in relatively well-defined regions of the cytoplasm. These results show that targeting and accumulation of the HIV Gag precursor, Pr55gag, at the plasma membrane occurs in yeast in the absence of other viral components and requires the N-myristoyl-glycine residue.
3. A novel monoclonal antibody to N-myristoyl glycine moiety found a new N-myristoylated HIV-1 p28gag protein in HIV-1-infected cells
K Furuishi, S Misumi, S Shoji Biochem Biophys Res Commun. 1996 May 15;222(2):344-51. doi: 10.1006/bbrc.1996.0746.
A novel monoclonal antibody was raised against a synthetic N-myristoyl glycine that is characteristic of all N-myristoylated proteins. The immunoreaction suppressed in the presence of hemocyanin as well as albumin conjugated with N-myristoyl glycine and other N-myristoyl glycyl peptides, while underivatized and myristoyl amino acid proteins or various fatty acids other myristic acid exerted no effect. The antibody specifically reacted with N-myristoylated pp60c-src in human colon adenocarcinoma cells, N-myristoylated pp60v-src in Rous sarcoma virus-infected cells, and N-myristoylated Gag precursor protein Pr55gag in HIV-1 producing cells. Furthermore, the antibody immunoreacted with a new N-myristoylated p28gag derived from HIV-1 gag protein. The antibody is shown to be a very useful tool for identification of N-myristoylated proteins.
