| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
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| Research Abstract |
Human semen-specific γ-glutamyl transpeptidase active protein (SEGAP) is an enzyme complex of ubiquitinated CD10 and CD13. In this study, we investigated the potential of SEGAP to induce tyrosine-phosrchorylation in HeLa (human cervical carcinoma), Raji (human B cell lymphoma) and HL60 (human promyelocytic leukemia). FACS analysis of SEGAP-treated cells revealed that SEGAP may bind to these cells. Western blots of SEGAP-and anti-CD13 antibody-treated cell lysates were performed using an anti-phosphotyrosine antibody. Phosphorylation of 120-kDa and 55-kDa proteins in HeLa and HL60 were observed in 1-min and 5-min incubations with an anti-CD13 antibody. Since CD13 is not distributed in Raji, 120-kDa and 55-kDa proteins were not phosphorylated. SEGAP phosphorylated 110-kDa proteins during 1-min incubation in HeLa and 110-kDa, 70-kDa and 60-kDa proteins during 1-min incubation in HL60. These results suggested that SEGAP induces tyrosine-phosphorylations of 110-kDa, 70-kDa and 60-kDa proteins which are thought to be as PI3 kinase, Syk/ZAP-70 and Src family proteins, respectively. Messenger RNA levels of cytokines, lectins and apotosis related-proteins were also examined in SEGAP-treated HL60. SEGAP did not affect mRNA levels of IL1-β, IL8, Lyn, Fas and TNF.INF-γ and mannose-binding lectin RNAs were not expressed in HL60, and their levels were not affected by SEGAP.
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