A NEW 185,000-DALTON SKELETAL-MUSCLE PROTEIN DETECTED BY MONOCLONAL-ANTIBODIES
A NEW 185,000-DALTON SKELETAL-MUSCLE PROTEIN DETECTED BY MONOCLONAL-ANTIBODIES
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DOI：
10.1083/jcb.98.2.518
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发表时间：
1984-01-01
影响因子：
7.8
通讯作者：
EPPENBERGER, HM
中科院分区：
文献类型：
--
作者：
GROVE, BK;KURER, V;EPPENBERGER, HM
M 线横穿[鸡]骨骼肌肌节粗丝区域的中心，似乎是多个结构元素的复杂阵列。迄今为止，有 2 种蛋白质与 M 系相关。它们是 MM-CK [meromyosin-肌酸激酶]，位于 M 4,4'' 亚纹中，以及 165,000 道尔顿 (164 kd) 蛋白，称为 M 蛋白和肌中蛋白。 185 kd 的第 3 个 M 系蛋白的阳性鉴定。在制备针对 165 kd 级分的单克隆抗体 (mAb) 的过程中，获得的 mAb 与分离的肌原纤维的 M 系结合（通过间接免疫荧光检测），但在以原始免疫原或低离子强度肌原纤维提取物作为抗原靶点的免疫印迹实验中识别出 185 kd 的蛋白条带。 185-kd 和 165-kd 蛋白质是不同蛋白质种类的证据是基于通过离子交换色谱将 2 种蛋白质分离成离散峰、其降解产物的独特模式以及 7 种 mAb 中任何一种的非交叉反应性。根据竞争性结合和免疫荧光实验确定，这些 mAb 可识别 185-kd 分子上的 3 个独特的抗原决定簇以及 165-kd 分子上的至少 2 个、可能是 4 个位点。为了解决165-kd蛋白质的多重命名问题，185-kd蛋白质将被称为肌中蛋白，而165-kd蛋白质将被称为M-蛋白质。
The M line, which transverses the center of the thick filament region of [chicken] skeletal muscle sarcomeres, appears to be a complex array of multiple structural elements. To date, 2 proteins are associated with the M line. They are MM-CK [meromyosin-creatine kinase], localized in the M 4,4'' substriations, and a 165,000-dalton (164 kd) protein, referred to as both M-protein and myomesin. The positive identification of a 3rd M-line protein of 185 kd. In the course of making monoclonal antibodies (mAb) against a 165-kd fraction, mAb were obtained that bound to the M line of isolated myofibrils as detected by indirect immunofluorescence, but recognized a protein band of 185 kd in immunoblotting experiments with either the original immunogen or low ionic strength myofibril extracts as antigenic targets. The evidence that the 185- and 165-kd proteins are distinct protein species is based on the separation of the 2 proteins into discrete peaks by ion exchange chromatograpy, the distinctive patterns of their degradation products, and non-cross-reactivity of any of 7 mAb. These mAb recognize 3 unique antigenic determinants on the 185-kd molecule and at least 2, and probably 4 sites on the 165-kd molecule as determined from competitive binding and immunofluorescence experiments. To resolve the problem of multiple nomenclature for the 165-kd protein, the 185-kd protein will be referred to a myomesin and the 165-kd protein as M-protein.