Ascaris suum:Protein Phosphotyrosine Phosphatases in Oocytes and Developing Stages

Wimmer, M., Schmid, B., Tag, C., and Hofer, H. W. 1998.Ascaris suum:Protein phosphotyrosine phosphatases in oocytes and developing stages.Experimental Parasitology88, 139–145. Protein tyrosine phosphatases were analyzed in oocytes ofAscaris suum. Phosphatases dephosphorylating modified acidic lysozyme were present in high-molecular-weight form (Mt>600,000) and as a 50-to 55-kDa protein in the soluble fraction. The low-molecular-weight form of the phosphatase cross-reacted with the antiserum raised against human T-cell protein tyrosine phosphatase and was not distinguishable from the 50- to 55-kDa protein tyrosine phosphatase previously described in the muscular layer of the adult worms (B. Schmidet al.1996,Molecular and Biochemical Parasitology77,183–192). The low-molecular-weight form was also present on immunoblots of high-molecular-weight protein tyrosine phosphatase preparations after denaturing electrophoresis. The same or a similar form of the tyrosine phosphatase was also found in detergent extracts from the pelletal fraction. In addition, another tyrosine phosphatase of 180 kDa molecular mass that dephosphorylated myelin basic protein was also found in extracts from the soluble compartment as well as detergent extracts from the pelletal fraction. It showed no cross-reactivity with antisera raised against soluble mammalian phosphatases and was resistant to inhibition by vanadate. While the activities of the myelin basic protein-dephosphorylating protein phosphatase remained fairly constant during early development of the oocytes, the activity of the enzyme dephosphorylating modified lysozyme in the pelletal fraction decreased to less than 10% of the initial activity between days 3 and 28 of incubation. Immuncytochemical studies of unfertilized and developingascariseggs revealed association of protein tyrosine kinase and protein tyrosine phosphatase with the egg shell, in addition to their presence in the neighborhood of mitochondria. The amount of enzyme changed with the stage of development. In the larval stage (21 days) protein tyrosine kinase had increased in the chitin layer of the shell and in the nuclei while the relative amount of tyrosine phosphatase decreased in accordance with the biochemical data.