Conformation-specific monoclonal antibodies directed against the calcium-stabilized structure of human prothrombin.
Conformation-specific monoclonal antibodies directed against the calcium-stabilized structure of human prothrombin.
复制标题
针对人凝血酶原的钙稳定结构的构象特异性单克隆抗体。
DOI：
10.1021/bi00273a037
复制
发表时间：
1983
期刊：
影响因子：
2.9
通讯作者：
Furie,B
中科院分区：
文献类型：
--
作者：
Lewis,RM;Furie,BC;Furie,B
Richard M. Lewis、Barbara C. Furie 和 Bruce Furie* 摘要：凝血酶原经历金属离子诱导的构象转变。为了制备针对人凝血酶原的金属稳定构象异构体的单克隆抗体，制备了鼠科体细胞杂交体，其精心设计了针对凝血酶原表面上的各种决定簇的抗体。鉴定了八个产生抗凝血酶原的克隆。所有八种抗体均结合人凝血酶原和凝血酶原片段 1，但未能结合前凝血酶 1、α-凝血酶或牛凝血酶原。四种单克隆抗体结合异常凝血酶原。这些抗体以比片段 1 更大的亲和力结合凝血酶原，强调了蛋白质及其片段之间的构象差异。一种抗体仅在存在 CaCl2 的情况下才能结合凝血酶原；该抗体不结合异常凝血酶原。鉴定了抗凝血酶原单克隆抗体的三个功能组：（1）不结合异常凝血酶原的金属稳定凝血酶原构象体特异性抗体；（2）结合凝血酶原和异常（脱7-羧基）pro-O^信息特异性抗体已被用作蛋白质结构探针，以补充化学和物理方法以确定蛋白质结构和功能的关系。免疫化学方法已应用于多肽构象运动的研究（Sachs 等，1972；Furie 等，1975；Hurrell 等，1977）、蛋白质折叠途径（Chavez 和 Scheraga，1977；Creighton 等）
Richard M. Lewis, Barbara C. Furie, and Bruce Furie* abstract: Prothrombin undergoes a conformational transition induced by metal ions. For preparation of monoclonal anti-bodies directed against the metal-stabilized conformer of hu-man prothrombin, murine somatic hybrids were prepared that elaborate antibodies against various determinants on the prothrombin surface. Eight anti-prothrombin-producing clones were characterized. All eight antibodies bound human prothrombin and prothrombin fragment 1 but failed to bind pre-thrombin 1, a-thrombin, or bovine prothrombin. Four mo-noclonal antibodies bound abnormal prothrombin. These antibodies bound prothrombin with greater affinity than fragment 1, emphasizing the conformational distinctions be-tween the protein and itsfragment. One antibody bound prothrombin only in the presence of CaCl2; this antibody did not bind abnormalprothrombin. Three functional groups of anti-prothrombin monoclonal antibodies were identified:(1) antibodies specific for the metal-stabilized prothrombin con-former that do not bind abnormalprothrombin;(2) antibodies that bind prothrombin and abnormal (des-7-carboxy) pro-O^ onformation-specific antibodies have been used as probes of protein structure to complement chemical and physical methods in determining the relationship of protein structure and function. Immunochemical approaches have been applied to the study of conformational motility of polypeptides (Sachs et al., 1972; Furie et al., 1975; Hurrell et al., 1977), pathways of protein folding (Chavez & Scheraga, 1977; Creighton et