人血红细胞酪氨酸蛋白磷酸酶(PTPP)的研究Ⅱ·胞浆PTPP的分离纯化及其主要性质

朱茂祥,吴国利

中国生物化学与分子生物学报 ›› 1994, Vol. 10 ›› Issue (02) : 174-179.

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PDF(372 KB)
中国生物化学与分子生物学报 ›› 1994, Vol. 10 ›› Issue (02) : 174-179.
论文

人血红细胞酪氨酸蛋白磷酸酶(PTPP)的研究Ⅱ·胞浆PTPP的分离纯化及其主要性质

  • 朱茂祥,吴国利
作者信息 +

Research for Phosphotyrosyl Protein Phosphatases (PTPPs) of Human Erythrocytes Ⅱ· Purification and Characterization of Cytosolic PTPP

  • Zhu,Mao-xiang;Wu,Guo-li
Author information +
文章历史 +

摘要

人血红细胞胞浆部分经(NH_4)_2SO_4沉淀,DEAE-纤维素(DE52)柱层析,磷酸纤维素柱层析(P11)得到部分纯化的PTPP,产率:5.7%,提纯1075倍。以(32) ̄P-Tyr-Poly(G_4:T)作底物,测得其表征Km约为0.5-0.8μmol/L,该酶的最适pH和最适温度分别为7.0-7.8及37-40℃。Zn ̄(2+)等二价金属离子及Na_3VO_4等酸根基团对其活性有明显的抑制作用;EDTA、甘油及DTT、巯基乙醇等则对其有强烈激活作用;而氟化物、酒石酸等对PTPP活性基本无影响。此外,某些蛋白质、氨基酸、核苷酸及抗肿瘤药物等对PTPP活性也都有不同程度的影响。特别是一些PKs及PPs在体外对PTPP活性也具有不同的作用。

Abstract

Through (NH_4)_2SO_4 partial precipitation, DEAE-cellulose chromatography and phospho-cellulose chromatography, PTPP can be purified partially from human erythrocytes cytosol with a 5.7% yield and a 1075-fold enrichment. Using ̄(32)P-Tyr-poly(G_4:T) as substrate, the apparent Km of the enzyme is 0. 5-0. 8μmol/L, and the optimum temperature and optimum pH are 37-40℃ and 7.0-7. 4, respectively. The PTPP activities can be inhibited by bivalent cations such as Zn ̄(2+) etc. and many anions(group) such as VO ̄(3-)_4 etc. EDTA,glycerol, DTT, mercaptoethanol etc. can stimulate the PTPP activities; Fluoride, tartrate,some inhibitor to acidic or alkaline phosphatases, have little effect on the PTPP activities. In addition, some proteins, nucleotides, amino acids, anti-tumour agents etc. have various effects on the PTPP activities. Especially, some PKs (protein kinases) and PPs (protein phosphatases) may regulate the PTPP activities, in vitro.

关键词

人血红细胞胞浆 / PTPP / 分离纯化 / 活性 / 抑制和激活

Key words

PTPP / Purification / Activity / Inhibition and stimulation

引用本文

导出引用
朱茂祥,吴国利. 人血红细胞酪氨酸蛋白磷酸酶(PTPP)的研究Ⅱ·胞浆PTPP的分离纯化及其主要性质[J]. 中国生物化学与分子生物学报, 1994, 10(02): 174-179
Zhu,Mao-xiang;Wu,Guo-li. Research for Phosphotyrosyl Protein Phosphatases (PTPPs) of Human Erythrocytes Ⅱ· Purification and Characterization of Cytosolic PTPP[J]. Chinese Journal of Biochemistry and Molecular Biology, 1994, 10(02): 174-179

参考文献

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