Protein S-myristoylation in Leishmania revealed with a heterologous reporter.

Reversible esterification of myristic acid to cysteine residue(s) (S-myristoylation) was documented recently in the protozoan Trypanosoma brucei. Unlike N-myristoylation, S-myristoylation appears to be rare (or non-existent) in animal cells and has not been documented in any other trypanosome. Reasoning that a lack of knowledge of appropriate substrates may have contributed to this state of affairs, we devised an assay to test for protein S-myristoylation in the ancient eukaryote Leishmania. A cDNA encoding a glycosylphosphatidylinositol-phospholipase C (GPI-PLC) from T. brucei was transfected into Leishmania and the expressed protein analyzed for covalent lipid modifications. Leishmania modified the reporter with myristate in a thio-ester linkage. From these observations, we infer that (i) GPI-PLC may be used as a reporter of this lipid modification in eukaryotes, and (ii) protein S-myristoylation might have ancient origins. Copyright 1999 Academic Press.