Identification of N-myristoylated proteins by reverse-phase high performance liquid chromatography of an azlactone derivative of N-myristoylglycine.
A method is presented for the identification of N-myristoylated proteins. N-Myristoyl transferases have an absolute requirement for a free N-terminal glycine. N-Myristoylglycine is released upon mild acid hydrolysis of myristoylated peptides and proteins and its derivitization to a p-nitrobenzylazlactone with subsequent analysis by reverse phase h.p.l.c. enabled its detection to pmol levels. This facilitated the identification of N-terminal myristate in nmol quantities of purified proteins. Using this method we demonstrate that the alpha-subunit of the GTP-binding protein G0 is N-terminally myristoylated.
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004151:Dipeptides; D019204:GTP-Binding Proteins; D005998:Glycine / Q000031:analogs & derivatives*; D006868:Hydrolysis; D019814:Myristic Acid; D009227:Myristic Acids / Q000032:analysis*; D011506:Proteins / Q000032:analysis*
D004151:Dipeptides; D009227:Myristic Acids; D011506:Proteins; D019814:Myristic Acid; C057393:N-myristoylglycine; D019204:GTP-Binding Proteins; D005998:Glycine