Park J W
Department of Biochemistry, College of Natural Sciences, Kyungpook National University, Korea.
Biochem Biophys Res Commun. 1996 Mar 7;220(1):31-5. doi: 10.1006/bbrc.1996.0351.
The effect of the S-nitrosothiol (RSNO) on the activation of NADPH oxidase in human neutrophils was studied using an in vitro translocation system in which an anionic amphiphil, such as sodium dodecyl sulfate or arachidonate, plays a role as an activator. When membranes pretreated with RSNO and a cytosol fraction from resting neutrophils were combined to reconstitute the NADPH oxidase, both translocation of the cytosolic NADPH oxidase components such as p47phox and p67phox to the plasma membrane fraction and subsequent superoxide generation was inhibited. However, RSNO had no effect on O2- production when added after enzyme activation. A similar inhibition of translocation of recombinant p47phox was observed with RSNO-treated membrane. When the RSNO-treated membrane fraction was exposed to 2-mercaptoethanol the inhibition was reversed. The data suggest that RSNO inhibits translocation of p47phox or p47phox containing cytosolic complex via a direct effect on the membrane component of the NADPH oxidase.
利用体外转位系统研究了S-亚硝基硫醇（RSNO）对人中性粒细胞中NADPH氧化酶激活的影响，在该系统中，阴离子两亲物如十二烷基硫酸钠或花生四烯酸盐作为激活剂发挥作用。当用RSNO预处理的膜与来自静息中性粒细胞的胞质溶胶部分结合以重建NADPH氧化酶时，胞质NADPH氧化酶成分如p47phox和p67phox向质膜部分的转位以及随后的超氧化物生成均受到抑制。然而，酶激活后添加RSNO对O2-产生没有影响。用RSNO处理的膜观察到重组p47phox转位的类似抑制。当RSNO处理的膜部分暴露于2-巯基乙醇时，抑制作用被逆转。数据表明，RSNO通过对NADPH氧化酶的膜成分的直接作用抑制p47phox或含p47phox的胞质复合物的转位。
