首页|Purification and Characterization of PRL Protein Tyrosine Phosphatases
Purification and Characterization of PRL Protein Tyrosine Phosphatases
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PRLs constitute a subfamily of protein tyrosine phosphatases(PTPs).In the present paper are reported the molecular cloning,expression,purification,and characterization of all the three members of the PRL en-zyrnes in human and the only PRL in C.elegans.These enzymes were expressed as glutathione S-transferase (GST) fusion proteins in DE3pLysS E.coli cells,and the recombinant fusion proteins were purified on glu-tathione-Sepharose affinity columns.Having been cleaved with thrombin,GST-free enzymes were further purified on an S-100 Sepharose gel filtration column.The purified proteins show single polypeptide bands on SDS-polyacrylamide gel electrophoresis.With para-nitrophenyl phosphate(p-NPP) as a substrate,PRLs exhibit classical Michaelis-Menten kinetics with V_(max) values two orders of magnitude smaller than those of classic PTPs.The responses of PRLs to ionic strength,metal ions and phosphatase inhibitors are similar to those of other characterized PTPs,but their optimal pH values are different.These data thus reveal distinct common biochemical properties of PRL subfamily PTPs as well.
PTPPRLPurificationCharacterization
LI Zhao-fa、WANG Yan、LI Qing-shan、ZHAO Zhi-zhuang Joe、FU Xue-qi
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Edmond H.Fischer Signal Transduction Laboratory,College of Life Sciences,Jilin University 9 Changchun 130023,P.R.China
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