Studies on the role of sulfhydryls in the myosin ATPase. Characterization of the site of modification by the bifunctional sulfhydryl reagent p-N,N'-phenylenedimaleimide.
Studies on the role of sulfhydryls in the myosin ATPase. Characterization of the site of modification by the bifunctional sulfhydryl reagent p-N,N'-phenylenedimaleimide.
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肌球蛋白 ATP 酶中巯基作用的研究。
DOI：
10.1016/s0021-9258(18)43503-x
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发表时间：
1980
期刊：
影响因子：
--
通讯作者：
P. J. Knight
中科院分区：
文献类型：
--
作者：
M. Burke;P. J. Knight
在MgADP存在的条件下，用接近化学计量量的双功能试剂[14C]p-N，N'-苯二马来酰亚胺（pPDM）修饰肌球蛋白，使其atp酶活性丧失。随后的羧甲基化和CNBr切割导致14C标签与Mr约10,000的单个多肽相关联。该肽的氨基酸组成和部分序列分析表明，它与Elzinga和Collins (Elzinga， M., and Collins， J.H. (1977) Proc. Natl测序的含有-SH1和-SH2的肽相对应。学会科学。由Kunz等人（Kunz, p.a., Walser， J.T, Watterson， J.G, and Schaub， M.C. (1977) FEBS Lett. 83, 137-140），和n -乙基马酰亚胺标记为-SH1或-SH2的肽。这些数据表明pPDM确实通过共价桥接来标记肌球蛋白中含有- sh1和- sh2的区域，并且表明在MgADP存在的情况下，这些硫醇可以接近12至14 A。
Myosin has been modified with near stoichiometric amounts of the bifunctional reagent [14C]p-N,N'-phenylenedimaleimide (pPDM) in the presence of MgADP under conditions which abolish its ATPase activity. Subsequent carboxymethylation and CNBr cleavage results in the 14C label being associated with a single polypeptide of Mr approximately 10,000. Amino acid composition and partial sequence analysis of this peptide showed that it corresponded to the peptide containing -SH1 and -SH2 sequenced by Elzinga and Collins (Elzinga, M., and Collins, J.H. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 4281-4284) and to the peptide labeled at -SH1 or -SH2 by N-ethylmaleimide by Kunz et al. (Kunz, P.A., Walser, J.T., Watterson, J.G., and Schaub, M.C. (1977) FEBS Lett. 83, 137-140). These data indicating that pPDM does label the -SH1- and -SH2-containing region in myosin by covalently bridging them and shows that in the presence of MgADP these thiols can approach to within 12 to 14 A.