F-actin is intermolecularly crosslinked by N,N'-p-phenylenedimaleimide through lysine-191 and cysteine-374.
F-actin is intermolecularly crosslinked by N,N'-p-phenylenedimaleimide through lysine-191 and cysteine-374.
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F-肌动蛋白通过 N,N-对亚苯基二酰亚胺通过赖氨酸 191 和半胱氨酸 374 进行分子间交联。
DOI：
10.1073/pnas.81.21.6599
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发表时间：
1984
影响因子：
11.1
通讯作者：
Phelan,JJ
中科院分区：
文献类型：
--
作者：
Elzinga,M;Phelan,JJ
双功能试剂 N,N'-p-亚苯基二酰亚胺 (PDM) 正用于测量 F-肌动蛋白内相邻单体中特定侧链之间的距离。合成[14C]PDM并用于交联F-肌动蛋白。通过凝胶过滤去除未交联的肌动蛋白，并且从共价交联二聚体和高级寡聚体的精氨酸特异性胰蛋白酶消化中，以高产率获得一种放射性交联肽。氨基酸组成和序列分析表明，它包含一个单体的残基 184-196 和相邻肌动蛋白分子的残基 373-375，通过 PDM 通过 Cys-374 和 Lys-191 桥接。因此，这些基团在 F-肌动蛋白中的相邻肌动蛋白单体中显示为相距 1.2-1.4 nm。该信息对于确定 F-肌动蛋白内肌动蛋白单体的方向可能至关重要。
The bifunctional reagent N,N'-p-phenylenedimaleimide (PDM) is being used in an attempt to measure distances between specific side chains in adjacent monomers within F-actin. [14C]PDM was synthesized and was used to crosslink F-actin. Uncrosslinked actin was removed by gel filtration, and, from an arginine-specific tryptic digest of the covalently crosslinked dimers and higher oligomers, one radioactive crosslinked peptide was obtained in high yield. Amino acid composition and sequence analysis indicated that it comprises residues 184-196 of one monomer and 373-375 of an adjacent actin molecule, bridged by PDM through Cys-374 and Lys-191. Thus, these groups are shown to be 1.2-1.4 nm apart in adjacent actin monomers in F-actin. This information may be crucial in establishing the orientation of actin monomers within F-actin.