Protein tyrosine phosphatase activity inLeishmania donovani

L. Donovani promastigotes were grown to late-log and 3-day stationary phase to determine the level of protein tyrosine phosphatase activity in crude extracts and in fractions following gel filtration column chromatography. Over 90% of the activity was soluble in a low salt extraction buffer in both phases of growth. Several peaks of activity were resolved following gel filtration of the crude extracts indicating that multiple tyrosine phosphatases are present in these cells. Tyrosine phosphatase activity was lower in 3-day stationary than in late log-phase cells and a reduction in the major peak of activity, eluting in a gel fraction corresponding to an M_r of approximately 168kDa, was observed.In vivo tyrosine phosphorylation was revealed by Western blot analysis. The degree of phosphorylation of at least two proteins differed in cells obtained from late log phase cultures as compared with 3-day stationary phase cultures. These observations indicate that changes in the balance between tyrosine phosphorylation and dephosphorylation occur with increasing culture age.Abbreviations MBP myelin basic protein - PMSF phenyl-methanesulfonylfluoride - PTP protein tyrosine phosphatase - RCML reduced, carboxyamidomethylated, maleylated lysozyme - YINAS Tyr-Ile-Asn-Ala-Ser