摘要
从人胎盘中分离得到谷胱甘肽S-转移酶纯化倍数470倍.比活力为37.897μmol/min·mg.经PAGE和SDS-PAGE证实为一条区带,亚基分子量为23000 Dalton;等电聚焦电泳结果,此酶等电点为4.6.底物GSH和CDNB的Km值分别为0.259mmol/L(r=0.97)和0.350mmol/L(r=0.99).不同浓度的巯基乙醇对GST-π活性有保护作用,1mmol/L浓度的保护作用较强.DEN,Triton X—100、苯巴比妥钠和AAF对GST-π活性起抑制作用,抑制作用随浓度增高而加强.
Glutathione S-transferase in Human placenta was purified appoximatly 470 fold by means of homogenate, ultracentrifuge, salting out, DEAE-cellulose (DE-52) column chromatography and GSH-sepharose 4B affinity chromatography. The specific activity of GST-π purified was 37.897U/mg, and SDS-PAGE, kinetic studies and isoelecctrofocusing were performed for the characterization of GST-π. The Km values of the GST-π for GSH and CDNB were 0.259 and 0.350 mmol/L, respectively. The molecular weight of subunit of GST-π was 23000 Dalton. Isoelectrofocusing of the enzyme showed only one band, and pI value was 4.6 and activities of GST-π were assayed treating with different concentration of mercapto ethanol,DEN,Triton X-100,sodium phenobarbital and AAF,respectively. The result showed that mercapto ethanol was a kind of protector, and the rest reagents were inhibitors for activity of GST-π.
出处
《延边医学院学报》
CAS
1992年第4期239-245,共7页
Journal of Medical Science Yanbian University
关键词
胎盘
谷胱甘肽转移酶类
酶试验
纯化
glutathione-transferases
enzyme tests
Placenta
purification
