Identification of two segments, separated by approximately 45 kilodaltons, of the myosin subfragment 1 heavy chain that can be cross-linked to the SH-1 thiol.
Identification of two segments, separated by approximately 45 kilodaltons, of the myosin subfragment 1 heavy chain that can be cross-linked to the SH-1 thiol.
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鉴定可与 SH-1 硫醇交联的肌球蛋白亚片段 1 重链的两个片段（相距约 45 千道尔顿）。
作者：
Sutoh,K;Lu,RC
材料和方法蛋白质和试剂。从家兔背部肌肉中提取肌球蛋白，并按Weeds和Pope（1977）的方法用糜蛋白酶消化制备SI。BPIA购自Molecular Probes（联合城，OR）。N ′-[7-（二甲基氨基）-4-甲基-3-香豆素基]马来酰亚胺（DACM）购自和子化学公司。（日本东京）。Vectastain和生物素化的抗兔IgG抗体购自Vector Laboratories（Burlingame，CA）。蛋白酶（金黄色葡萄球菌，菌株V8）、胰凝乳蛋白酶和赖氨酰内肽酶购自Miles Scientific（Na-perville，IL）、Sigma（St. Louis，MO）和和子Chemical Co.（日本东京）。针对肌球蛋白重链N-末端的多克隆抗体如下产生。通过固相法合成具有序列Ac-Ser-Ser-Asp-Ala-Asp-Met-Ala-Val-Lys的肽（Stewart，1969）。该肽的序列对应于肌球蛋白重链N-末端区域的前8个残基（Starr & Offer，1973; Tong & Elzinga，1984），除了在C-末端添加赖氨酸残基。通过戊二醛将匙孔冲击血蓝蛋白（KLH）偶联至合成肽的赖氨酸残基（Bulinski等人，1983年）。KLH-肽复合物中的表位密度为10。如前所述在新西兰白色兔中产生抗体（Bulinski等人，1983年）。用酶联免疫吸附试验（ELI-SA）测得抗体滴度为105。肽合成和抗体表征的细节将在别处公开（Sutoh et al.，1987年）。BPIA对SI的改性及光解研究SH-1在缓冲液中以1：1.5摩尔比的SI与BPIA进行修饰
Materials and MethodsProteins and Reagents. Myosin was prepared fromrabbit back muscle, and SI was made by chymotrypsin digestion according to Weeds andPope (1977). BPIA was purchased from Molecular Probes (Junction City, OR). A'-[7-(Dimethylamino)-4-methyl-3-coumarinyl] maleimide (DACM) was from Wako Chemical Co.(Tokyo, Japan). Vectastain and a biotinylated anti-rabbit IgG antibody were purchased from Vector Laboratories (Burlingame, CA). Protease {Staphylococcus aureus, strain V8), chymotrypsin, and lysyl endopeptidase were purchased from Miles Scientific (Na-perville, IL), Sigma (St. Louis, MO), and Wako Chemical Co.(Tokyo, Japan), respectively. A polyclonal antibody against the N-terminus of the myosin heavy chain was raised as follows. A peptide with the sequence Ac-Ser-Ser-Asp-Ala-Asp-Met-Ala-Val-Lys was synthesized by the solid-phase method (Stewart, 1969). The sequence of this peptide corresponds to the first eight residues of the N-terminal region of the myosin heavy chain (Starr & Offer, 1973; Tong & Elzinga, 1984) except that a lysine residue was added at the C-terminus. Keyholelimpet hemocyanin (KLH) was coupled to the lysine residue of the synthetic peptide by glutaraldehyde (Bulinski et al., 1983). The epitope density in the KLH-peptide complex was 10. Antibody was raised in New Zealand White rabbits as previously described (Bulinski et al., 1983). The titer of the antibody was estimated to be 105 by the enzyme-linked immunosorbant assay (ELI-SA). Details of the peptide synthesis and the characterization of the antibody will be published elsewhere (Sutoh et al., 1987). Modification and Photolysis of SI with BPIA. SH-1 was modified at a 1: 1.5 molar ratio of SI to BPIA in a buffer
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使用荧光交联剂稳定肌球蛋白亚片段 1 中的初级环。
DOI：
10.1073/pnas.82.6.1658
发表时间：
1985
影响因子：
11.1
作者：
Mornet,D;Ue,K;Morales,MF
通讯作者：
Morales,MF
通过蛋白水解酶消化揭示肌球蛋白亚片段 1 的亚结构。
影响因子：
2.7
作者：
K. Yamamoto;T. Sekine
通讯作者：
T. Sekine
DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Perkins,WJ;Weiel,J;Grammer,J;Yount,RG
通讯作者：
Yount,RG
骨骼肌细丝和肌球蛋白分子复合物的三维图像分析。
DOI：
--
发表时间：
1985
期刊：
Journal of Biochemistry (Tokyo)
影响因子：
--
作者：
C. Toyoshima;T. Wakabayashi
通讯作者：
T. Wakabayashi
N-乙基马来酰亚胺修饰肌球蛋白的酶学特性
DOI：
--
发表时间：
1964
期刊：
影响因子：
--
作者：
T. Sekine;W. W. Kielley
通讯作者：
W. W. Kielley