Eaton B L, Pepe F A
J Cell Biol. 1972 Dec;55(3):681-95. doi: 10.1083/jcb.55.3.681.
M band protein can be specifically extracted from fresh chicken breast muscle myofibrils suspended in 5 mM Tris-HCl pH 8.0. During discontinuous polyacrylamide gel electrophoresis the isolated protein separates into three bands which can be identified as two separate components (A, B) and a complex of the two. When partially purified fractions of the separated components are combined, an increase in the intensity of the band containing the complex can be shown. The polypeptide chain weights of the two components are 100,000 (A) and 40,000 (B) daltons as estimated by sodium dodecyl sulfate- (SDS-) polyacrylamide gel electrophoresis. Antibody prepared against total M band protein stains only the M band of the myofibril and is completely absorbed by M band protein. M band protein also absorbs the M band staining specifically from antibody which stains both I and M bands. Immunodiffusion data indicate that anti-M band is a mixture of two specific antibodies, one against each component.
M带蛋白可从悬浮于5 mM Tris-HCl(pH 8.0)中的新鲜鸡胸肌肌原纤维中特异性提取。在不连续聚丙烯酰胺凝胶电泳过程中,分离出的蛋白可分为三条带,这三条带可被鉴定为两个单独的组分(A、B)以及二者的复合物。当将分离组分的部分纯化级分混合时,含有复合物的条带强度会增加。通过十二烷基硫酸钠-(SDS-)聚丙烯酰胺凝胶电泳估计,这两个组分的多肽链分子量分别为100,000(A)和40,000(B)道尔顿。针对总M带蛋白制备的抗体仅对肌原纤维的M带染色,并且会被M带蛋白完全吸收。M带蛋白还能特异性地从同时对I带和M带染色的抗体中吸收M带染色。免疫扩散数据表明,抗M带是两种特异性抗体的混合物,每种抗体针对一个组分。