W Tumor proliferation of cancer cells requires a high intake of oxygen by angiogenesis. Deep cancer cells suffer from asphyxia and meet their energy needs through the enzymes of glycolysis. The anti-angio- genesis app...W Tumor proliferation of cancer cells requires a high intake of oxygen by angiogenesis. Deep cancer cells suffer from asphyxia and meet their energy needs through the enzymes of glycolysis. The anti-angio- genesis approach has been recognized for therapeutic purposes, but the deep cancers, difficult to reach by this therapy, could be targeted by inhibiting an enzyme of the glycolytic cycle. Our work focused on the study of the expression of GAPDH, a key enzyme of glycolysis, in cervix, breast and prostate tumors, for two approaches: Fundamental and targeted therapeutics. 60 samples, taken at the Anatomopathology laboratory of the Pasteur Institute of Morocco, were examined histologically and immunohistochemically, demonstrating the expression and cellular localization of GAPDH. The three organs have shown an overex-pression of GAPDH in tumor tissues. At the cellular level, the localization of GAPDH in cancer tissue is diffuse but mostly nuclear whereas it remains focused at the membrane and/or the cytoplasm in benign tumor tissues. From these results we could assume that GAPDH is involved in the cancer process and draws attention to a possible new nuclear role that could be either specific to one form or different isoforms of GAPDH enzyme.展开更多
Idiomarina loihiensis was isolated from the salt works in Sfax (Tunisia), until now, the characterization of the GAPDH phosphorylante was never studied. Here, we report the isolation and the biochemical characterizati...Idiomarina loihiensis was isolated from the salt works in Sfax (Tunisia), until now, the characterization of the GAPDH phosphorylante was never studied. Here, we report the isolation and the biochemical characterization of glyceralehyde-3-phosphate dehydrogenase (GAPDH) fromI. loihiensis saline’s bacteria on the basis of the apparent native and subunit molecular weights, physico-chemical and kinetic characterizations. The purification method consisted of two steps, ammonium sulfate fractionation followed by one chromatographic step, namely dye-affinity on Blue Sepharose CL-6B. Polyclonal antibodies against the purified enzyme were used to recognize theI. loihiensis GAPDH by Western blotting. The optimum pH of the purified enzyme was 8.5. Studies on the effect of temperatures revealed an enzyme increasing activity of about 45?C. The molecular weight of the purified enzyme was 36 kDa determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Non-denaturing polyacrylamide gels yield a molecular weight of 147 kDa. The Michaelis constants for NAD+ and D-glyceraldehyde-3-phosphate estimated was 19 μM and 3.1 μM, respectively. The maximal velocity of the purified enzyme was estimated to be 2.06 U/mg, approximately 6-fold increase in specific activity and a final yield of approximately 32.5%. The physicochemical properties of this GAPDH, being characterized, could be used in further studies.展开更多
文摘W Tumor proliferation of cancer cells requires a high intake of oxygen by angiogenesis. Deep cancer cells suffer from asphyxia and meet their energy needs through the enzymes of glycolysis. The anti-angio- genesis approach has been recognized for therapeutic purposes, but the deep cancers, difficult to reach by this therapy, could be targeted by inhibiting an enzyme of the glycolytic cycle. Our work focused on the study of the expression of GAPDH, a key enzyme of glycolysis, in cervix, breast and prostate tumors, for two approaches: Fundamental and targeted therapeutics. 60 samples, taken at the Anatomopathology laboratory of the Pasteur Institute of Morocco, were examined histologically and immunohistochemically, demonstrating the expression and cellular localization of GAPDH. The three organs have shown an overex-pression of GAPDH in tumor tissues. At the cellular level, the localization of GAPDH in cancer tissue is diffuse but mostly nuclear whereas it remains focused at the membrane and/or the cytoplasm in benign tumor tissues. From these results we could assume that GAPDH is involved in the cancer process and draws attention to a possible new nuclear role that could be either specific to one form or different isoforms of GAPDH enzyme.
文摘Idiomarina loihiensis was isolated from the salt works in Sfax (Tunisia), until now, the characterization of the GAPDH phosphorylante was never studied. Here, we report the isolation and the biochemical characterization of glyceralehyde-3-phosphate dehydrogenase (GAPDH) fromI. loihiensis saline’s bacteria on the basis of the apparent native and subunit molecular weights, physico-chemical and kinetic characterizations. The purification method consisted of two steps, ammonium sulfate fractionation followed by one chromatographic step, namely dye-affinity on Blue Sepharose CL-6B. Polyclonal antibodies against the purified enzyme were used to recognize theI. loihiensis GAPDH by Western blotting. The optimum pH of the purified enzyme was 8.5. Studies on the effect of temperatures revealed an enzyme increasing activity of about 45?C. The molecular weight of the purified enzyme was 36 kDa determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Non-denaturing polyacrylamide gels yield a molecular weight of 147 kDa. The Michaelis constants for NAD+ and D-glyceraldehyde-3-phosphate estimated was 19 μM and 3.1 μM, respectively. The maximal velocity of the purified enzyme was estimated to be 2.06 U/mg, approximately 6-fold increase in specific activity and a final yield of approximately 32.5%. The physicochemical properties of this GAPDH, being characterized, could be used in further studies.
