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Characterization of a tyrosine phosphatase activity in the oogenesis of Periplaneta americana
Author(s) -
Oliveira D.M.P.,
Machado E.A.
Publication year - 2006
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.20139
Subject(s) - acid phosphatase , phosphatase , biology , biochemistry , yolk , alkaline phosphatase , sodium orthovanadate , protein tyrosine phosphatase , phosphoserine , microbiology and biotechnology , tyrosine , enzyme , ecology , serine
Abstract In this work, phosphatase activity was characterized in the ovary and the haemolymph of Periplaneta americana . The optimum pH for these activities was 4.0, and a temperature of 44°C was ideal for the maximal enzyme activity. The phosphatase activities were inhibited by NaF, sodium tartrate, Pi, sodium orthovanadate, and ammonium molybdate. The ovarian phosphatase activity at pH 4.0 was almost exclusive against phosphotyrosine, with little or no effect on the residues of phosphoserine or phosphothreonine. These results indicate that this phosphatase activity is due to the presence of an acid tyrosine phosphatase. The phosphatase activities of acid extracts from P. americana ovaries (OEX) and an acid extract from P. americana haemolymph (HEX) were analyzed in non‐denaturant gel electrophoresis using an analog substrate β‐naphtyl phosphate. The gel revealed two bands with phosphatase activity in the ovary and one band in the haemolymph; these bands were excised and submitted to a 10% SDS‐PAGE showing a single 70‐kDa polypeptide in both samples. Histochemistry of the ovary with α‐naphtyl phosphate for localization of acid phosphatase activity showed mainly labeling associated to the oocyte peripheral vesicles, basal lamina, and between follicle cells. Electron microscopy analysis showed that acid phosphatase was localized in small peripheral vesicles in the oocyte, but not inside yolk granules. The possible role of this phosphatase during oogenesis and embryogenesis is also discussed in this article. Arch Insect Biochem Physiol 63:24–35, 2006. © 2006 Wiley‐Liss, Inc.