Klinger E, Aviram I
Department of Biochemistry, Faculty of Life Sciences, Tel-Aviv University, Israel.
Biochem Biophys Res Commun. 1991 May 31;177(1):504-10. doi: 10.1016/0006-291x(91)92012-9.
Superoxide production by neutrophil NADPH oxidase activated in a cell-free system consisting of plasma membranes, cytosol and arachidonate is enhanced by nonhydrolyzable analogs of GTP and reduced by GDP. To characterize the interaction of guanine nucleotides with the system, dialdehyde analogs of GTP and GDP (oGTP and oGDP) were employed. oGDP or oGTP caused an irreversible and dose dependent inactivation of NADPH oxidase-supporting cytosolic activity. Cytosol was fractionated on S and Q Sepharose ion exchange columns into three fractions, combinations of which synergistically supported activation of NADPH oxidase. Two fractions shown by immunoblotting to contain the oxidase-linked p47 and p67 proteins were inactivated by oGDP. Labeling with [alpha-32P]-oGTP lead to incorporation of the label into several proteins.
在由质膜、胞质溶胶和花生四烯酸组成的无细胞系统中被激活的中性粒细胞NADPH氧化酶产生超氧化物的过程，会被GTP的不可水解类似物增强，被GDP降低。为了表征鸟嘌呤核苷酸与该系统的相互作用，使用了GTP和GDP的二醛类似物（oGTP和oGDP）。oGDP或oGTP导致NADPH氧化酶支持的胞质活性发生不可逆的剂量依赖性失活。胞质溶胶在S和Q Sepharose离子交换柱上被分离成三个部分，这些部分的组合协同支持NADPH氧化酶的激活。通过免疫印迹显示含有氧化酶相关p47和p67蛋白的两个部分被oGDP失活。用[α-32P]-oGTP标记导致该标记掺入几种蛋白质中。
