NAD-dependent cross-linking of dinitrogenase reductase and dinitrogenase reductase ADP-ribosyltransferase from Rhodospirillum rubrum.
NAD-dependent cross-linking of dinitrogenase reductase and dinitrogenase reductase ADP-ribosyltransferase from Rhodospirillum rubrum.
复制标题
红色红螺菌二固氮酶还原酶和二固氮酶还原酶 ADP-核糖基转移酶的 NAD 依赖性交联。
DOI：
10.1128/jb.179.10.3277-3283.1997
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发表时间：
1997
影响因子：
3.2
通讯作者：
Ludden,PW
中科院分区：
文献类型：
--
作者：
Grunwald,SK;Ludden,PW
用1-乙基-3-（3-二甲氨基丙基）-碳二亚胺和磺基-N-羟基丁二酰亚胺两种交联剂，研究了红色红酵母双固氮酶还原酶和双固氮酶还原酶ADP-核糖基转移酶（DRAT）的化学交联。二固氮酶还原酶和DRAT之间的交联需要存在NAD、细胞ADP-核糖供体或含有未修饰的烟酰胺基团的NAD类似物，如烟酰胺次黄嘌呤二核苷酸。NADP在修饰反应中不会取代NAD，但确实支持二氮还原酶和DRAT之间的交联。DRAT催化的ADP-核糖基化的二氮酶还原酶抑制氯化钠，是二氮酶还原酶和DRAT之间的交联，这表明离子相互作用所需的这两种蛋白质的协会。交联对于天然的、未修饰的二氮酶还原酶是特异性的，因为氧变性的和ADP-核糖基化的二氮酶还原酶都不能与DRAT形成交联复合物。二氮酶还原酶的ADP结合和腺嘌呤核苷酸游离状态与DRAT形成交联复合物;然而，当二氮酶还原酶处于其ATP结合状态时，交联被抑制。
Chemical cross-linking of dinitrogenase reductase and dinitrogenase reductase ADP-ribosyltransferase (DRAT) from Rhodospirillum rubrum has been investigated with a cross-linking system utilizing two reagents, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and sulfo-N-hydroxysuccinimide. Cross-linking between dinitrogenase reductase and DRAT requires the presence of NAD, the cellular ADP-ribose donor, or a NAD analog containing an unmodified nicotinamide group, such as nicotinamide hypoxanthine dinucleotide. NADP, which will not replace NAD in the modification reaction, does support cross-linking between dinitrogenase reductase and DRAT. The DRAT-catalyzed ADP-ribosylation of dinitrogenase reductase is inhibited by sodium chloride, as is the cross-linking between dinitrogenase reductase and DRAT, suggesting that ionic interactions are required for the association of these two proteins. Cross-linking is specific for native, unmodified dinitrogenase reductase, in that both oxygen-denatured and ADP-ribosylated dinitrogenase reductase fail to form a cross-linked complex with DRAT. The ADP-bound and adenine nucleotide-free states of dinitrogenase reductase form cross-linked complexes with DRAT; however, cross-linking is inhibited when dinitrogenase reductase is in its ATP-bound state.