Chemical characterization of p17gag from human immunodeficiency virus as an N-terminally myristoylated protein.
The N-terminal p17gag protein of the human immunodeficiency virus has been shown to incorporate radioactivity following labelling of infected cell lines with [3H]myristic acid. We investigated p17gag to determine whether the incorporated radioactivity was the consequence of N-terminal myristoylation. The virus was purified by density gradient centrifugation after labelling chronically infected H9 cells with [3H]myristic acid. The p17gag was isolated by immunoprecipitation and subjected to partial acid hydrolysis. [3H]Myristoylglycine generated by the hydrolysis was derivatized to 4-(p-nitrobenzylidene)-2-tridecanoyloxazol-5-one and identified against a co-eluting, derivatized, unlabelled N-myristoylglycine standard by reverse-phase high-performance liquid chromatography. This study unequivocally demonstrates that p17gag is an N-myristoylated protein.
Goddard
C
C
Division of Cancer Treatment, National Cancer Institute, Bethesda, MD 20892.
Aquino
A
A
Glazer
R I
RI
Felsted
R L
RL
D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D015683:Gene Products, gag; D006678:HIV / Q000032:analysis* / Q000302:isolation & purification; D015488:HIV Antigens / Q000032:analysis*; D006801:Humans; D007553:Isotope Labeling; D019814:Myristic Acid; D009227:Myristic Acids / Q000032:analysis* / Q000378:metabolism; D010443:Peptide Chain Termination, Translational; D011233:Precipitin Tests; D012191:Retroviridae Proteins / Q000032:analysis*; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D015488:HIV Antigens; D009227:Myristic Acids; D012191:Retroviridae Proteins; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C054858:p17 protein, Human Immunodeficiency Virus Type 1; D019814:Myristic Acid