Körner M, Van Thiem N, Cardinaud R, Lacombe G
Biochemistry. 1983 Dec 6;22(25):5843-7. doi: 10.1021/bi00294a024.
Cardiac and skeletal myosin subfragments 1 cleaved into three fragments were modified by 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide metho-p-toluene-sulfonate in the presence of the nucleophile nitrotyrosine ethyl ester. The effects observed (first-order kinetics of ATPase inactivation, incorporation of 1 mol of nitrotyrosine/mol of subfragment 1) were similar to those previously observed for the nondigested subfragments 1 [Lacombe, G., Van Thiem, N., & Swynghedauw, B. (1981) Biochemistry 20, 3648-3653; Körner, M., Van Thiem, N., Lacombe, G., & Swynghedauw, B. (1982) Biochem. Biophys. Res. Commun. 105, 1198-1207]. For both native and digested subfragments 1, which were inactivated to the extent of about 70%, the location of the label nitrotyrosine was performed by immunological blotting with 125I-labeled anti-nitrotyrosine immunoglobulins. It was found that the modified residue was essentially located on the heavy chain for the native subfragments 1 and on the 50K peptide for the digested subfragments 1.
将心肌和骨骼肌肌球蛋白亚片段1切割成三个片段，在亲核试剂硝基酪氨酸乙酯存在的情况下，用1-环己基-3-(2-吗啉乙基)碳二亚胺对甲苯磺酸盐进行修饰。观察到的效应（ATP酶失活的一级动力学，每摩尔亚片段1掺入1摩尔硝基酪氨酸）与先前对未消化的亚片段1观察到的效应相似[拉孔布，G.，范蒂姆，N.，& 斯文赫道，B.（1981年）《生物化学》20，3648 - 3653；克纳，M.，范蒂姆，N.，拉孔布，G.，& 斯文赫道，B.（1982年）《生物化学与生物物理研究通讯》105，1198 - 1207]。对于天然和消化后的亚片段1，其失活程度约为70%，通过用125I标记的抗硝基酪氨酸免疫球蛋白进行免疫印迹来确定硝基酪氨酸标记的位置。结果发现，修饰的残基在天然亚片段1中主要位于重链上，在消化后的亚片段1中位于50K肽上。
