The myosin SH2-50-kilodalton fragment cross-link: location and consequences.
The myosin SH2-50-kilodalton fragment cross-link: location and consequences.
复制标题
肌球蛋白 SH2-50 千道尔顿片段交联：位置和后果。
DOI：
10.1021/bi00405a059
复制
发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Kassab,R
中科院分区：
文献类型：
--
作者：
Chaussepied,P;Morales,MF;Kassab,R
材料与方法材料。A-糜蛋白酶来自沃辛顿生化公司。A，A“-对亚苯二马来酰亚胺来自伯林格-曼海因。溴化氰和EDC来自Sigma。[14C]ADP来自新英格兰核。Na-DodS04、丙烯酰胺和所有其他凝胶电泳剂都来自Bio-Rad。A-乙基吗啉、丹磺酰氯、二甲基甲酰胺和F-1700微聚酰胺平板均来自皮尔斯。所有其他化学品的分析等级都是最高的。蛋白质及其硫代衍生物的制备。按照Offer等人(1973)的描述制备兔骨骼肌球蛋白。SL是通过消化肌球蛋白细丝1得到的缩写：SL，肌球蛋白胰凝乳酶亚段1；LC 1，碱性轻链1；LC 3，碱性轻链3；SHJ，硫醇残基707；SH2，硫醇残基697；MR，相对分子质量；CD，圆二色；PPDM，N，-N‘-对苯二马来酰亚胺；DNFB，2，4-二硝基氟苯；DNP-S-1，二硝基苯基亚段1；IAA，碘乙酰胺；NEM，TV-乙基雄酰亚胺；IAEDANS，N-(碘乙酰基)-7V’(5-磺基-L-萘基)乙二亚胺；DTE，二硫代乙二酚；EDC，1-乙基-3-[3-(二甲氨基)-丙基]碳二亚胺；NaDODS-04，十二烷基硫酸钠；HEPES，N-(2-羟乙基)哌嗪-Ar‘-2-乙磺酸；三-盐酸盐，三羟甲基)氨基甲烷盐酸盐；MOPS，4-吗啉丙磺酸；AEDANS，7V-acetyl-lV’-(5-sulfo-l-naphthyl)-ethylenediamine；kDa，千道尔顿(S)；乙二胺四乙酸。
Materials and Methods Materials. a-Chymotrypsin was from Worthington Bio-chemicals. A, A"-p-Phenylenedimaleimide was from Boeh-ringer-Mannhein. Cyanogen bromide and EDC were from Sigma.[14C] ADP was from New England Nuclear. Na-DodS04, acrylamide, and all other gel electrophoresis chemicals were from Bio-Rad. A-Ethylmorpholine, dansyl chloride, dimethylformamide, and F-1700 micro-polyamide plates were from Pierce. All other chemicals were of the highest analytical grade. Preparation of Proteins and Sl Derivatives. Rabbit skeletal myosin was prepared as described by Offer et al.(1973). Sl was obtained by digestion of myosin filaments1 Abbreviations: Sl, myosin chymotryptic subfragment 1; LC1, alkali light chain 1; LC3, alkali light chain 3; SHj, thiol residue 707; SH2, thiol residue 697; Mr, molecular weight; CD, circular dichroism; pPDM, N,-N'-p-phenylenedimaleimide; DNFB, 2, 4-dinitrofluorobenzene; DNP-S-1, dinitrophenyl subfragment 1; IAA, iodoacetamide; NEM, TV-ethylmale-imide; IAEDANS, N-(iodoacetyl)-7V'-(5-sulfo-l-naphthyl) ethylenediamine; DTE, dithioerythritol; EDC, 1-ethyl-3-[3-(dimethylamino)-propyl] carbodiimide; NaDodS04, sodium dodecyl sulfate; Hepes, N-(2-hydroxyethyl) piperazine-Ar'-2-ethanesulfonic acid; Tris-HCl, trisfhydroxymethyl) aminomethane hydrochloride; Mops, 4-morpholinepropanesulfonic acid; AEDANS, 7V-acetyl-lV'-(5-sulfo-l-naphthyl)-ethylenediamine; kDa, kilodalton (s); EDTA, ethylenediaminetetraacetic acid.