Tunwell R E, O'Connor C D, Mata A M, East J M, Lee A G
Department of Biochemistry, University of Southampton, U.K.
Biochim Biophys Acta. 1991 Apr 9;1073(3):585-92. doi: 10.1016/0304-4165(91)90234-8.
Epitopes for a number of monoclonal antibodies (mAbs) binding (Ca(2+)-Mg2+)-ATPase purified from skeletal muscle sarcoplasmic reticulum have been defined by studying binding to fusion proteins generated from cDNA fragment libraries. Comparison of these results with those of previous studies of binding of mAbs to proteolytic fragments of the ATPase have allowed the definition of the epitopes to within approx. 100 residues and for one (mAb 1/2H7) to within 45 residues. The experiments suggest considerable exposure of the nucleotide binding domain of the ATPase on the top surface of the protein. Those mAbs that were found to inhibit steady-state ATPase activity were found to bind to epitopes in the nucleotide binding domain of the ATPase.
通过研究从骨骼肌肌浆网纯化的（Ca²⁺-Mg²⁺）-ATP酶与cDNA片段文库产生的融合蛋白的结合，已确定了多种单克隆抗体（mAb）的表位。将这些结果与先前关于mAb与ATP酶蛋白水解片段结合的研究结果进行比较，已将表位定义在约100个残基范围内，对于一种抗体（mAb 1/2H7），已定义在45个残基范围内。实验表明，ATP酶的核苷酸结合结构域在蛋白质的顶表面上有相当大的暴露。那些被发现抑制稳态ATP酶活性的mAb被发现与ATP酶核苷酸结合结构域中的表位结合。
