Identification and purification of membrane and soluble forms of the major surface protein of Leishmania promastigotes.

A major integral membrane glycoprotein of 63 kDa (p63), present at 500,000 copies/cell, was found on the surface of Leishmania major LEM 513 promastigotes. This protein was labeled either by surface iodination of the cells or by metabolic incorporation of [35S]methionine. Peptide maps of the proteins labeled by the two procedures were identical. Protein p63 was purified in three steps: extraction and phase separation in the nonionic detergent Triton X-114, chromatography on DEAE-cellulose, and finally chromatography on a Mono-Q column. The carbohydrate content as well as the concanavalin A receptor activity were characterized. A hydrophilic form of p63 was generated during the purification of the protein. This form was not derived by proteolysis from the amphiphilic protein found in the membrane, but may have been generated by the hydrolysis of a lipid containing myristyl residue(s) anchoring the protein in the membrane.