摘要
用免疫亲和层析结合常规生化方法从人胸腺中纯化腺苷脱氨酶达电泳纯，比活力１４８９８Ｕ／ｍｇ，得率３４．１５％该酶分子质量为４１．３ｋｕ，约由３８０个氨基酸残基构成，等电点为４．９，最适温度３７～４０℃，最适ｐＨ为７．０以腺苷或２－脱氧腺苷为底物，其Ｋｍ分别为８３μｍｏｌ／Ｌ和６１μｍｏｌ／Ｌ．对氯汞苯甲酸能显著抑制酶活性，二硫苏糖醇可使被抑制的活性得到部分恢复．
A simplified procedure for the purification of adenosine deaminase(ADA)fromhuman thymus based on immuno-affinity chromatography of anti-human thymus ADA IgGwas described. After ammonium sulfate fractionation;immuno-affinity chromatography andSephadex G-100 gel filtration. ADA was separated as homogeneity from human thymus.Theyield and the specific activity of purified ADAwere 34.15% and 14898 U/mg respectively.The purified ADA molecule consists of about 380amino acid residues giving a Mr of 41. 3 ku andPI of 4.9.The optimum temperature is 37～40℃. The optimum PH is 7 .0. Using adenosineor 2-deoxyadenosine as substrate the apparentKm of the enzyme is 83 μmol/L and 61 μmol/Lrespectively.The enzyme activity can beinhibited by p-chloromercuric benzoic acid whilepartially restored by dithiothreitol. ADA activitywas decreased by anti-calf ADA IgG and antithymus ADA IgG.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1996年第6期531-537,共7页
Progress In Biochemistry and Biophysics
基金
江西省自然科学基金
关键词
腺苷脱氨酶
免疫亲和纯化
性质
adenosine deaminase
immuno affinity chromatography
property
human thymus
