Ogata K, Ogata Y, Takasaki Y, Tan E M
W.M. Keck Autoimmune Disease Center, Department of Basic & Clinical Research, Scripps Clinic and Research Foundation, La Jolla, CA 92037.
J Immunol. 1987 Nov 1;139(9):2942-6.
The immune epitopes of proliferating cell nuclear antigen (PCNA), also called cyclin, were analyzed by determining the reactivity between PCNA peptide fragments and anti-PCNA antibodies from lupus patients, murine monoclonal antibody (19A2), and rabbit anti-NH2-terminal peptide antibody. Limited digestion of PCNA/cyclin with Staphylococcus aureus V8 protease resulted in several peptide fragments. Five fragments of 30, 20, 15, 14, and 13 kDa were reactive with rabbit anti-NH2-terminal peptide antibody denoting that they contained the NH2-terminal peptide. The 30- and 20-kDa fragments reacted with 19A2 but the others did not. Lupus sera reacted with 17- and 15-kDa peptide fragments allowing their classification into three groups. Two of eight sera (type A) reacted only with the 17-kDa fragment. Two others (type B) reacted with both the 17- and 15-kDa fragments and the remaining four sera (type C) reacted only with the 15-kDa fragment. The sera reacting with the 15-kDa fragment also reacted with the 20-kDa fragment, but the sera reactive only with the 17-kDa fragment did not, indicating that the 17-kDa fragment was not a degradation product of 20-kDa fragments. The 19A2 epitope resided in the region between 15 and 20 kDa from the NH2 terminus, whereas there was at least one distinct epitope on each 15- and 17-kDa peptide, which were recognized by lupus autoantibodies.
通过测定增殖细胞核抗原(PCNA,又称细胞周期蛋白)肽段与狼疮患者的抗PCNA抗体、鼠单克隆抗体(19A2)以及兔抗NH2末端肽抗体之间的反应性,对PCNA的免疫表位进行了分析。用金黄色葡萄球菌V8蛋白酶对PCNA/细胞周期蛋白进行有限消化,产生了几个肽段。5个分别为30、20、15、14和13 kDa的肽段与兔抗NH2末端肽抗体发生反应,表明它们含有NH2末端肽。30 kDa和20 kDa的肽段与19A2发生反应,但其他肽段不反应。狼疮血清与17 kDa和15 kDa的肽段发生反应,可将其分为三组。8份血清中的2份(A型)仅与17 kDa的肽段发生反应。另外2份(B型)与17 kDa和15 kDa的肽段均发生反应,其余4份血清(C型)仅与15 kDa的肽段发生反应。与15 kDa肽段发生反应的血清也与20 kDa的肽段发生反应,但仅与17 kDa肽段发生反应的血清则不然,这表明17 kDa的肽段不是20 kDa肽段的降解产物。19A2表位位于从NH2末端起15至20 kDa的区域内,而在15 kDa和17 kDa的每个肽段上至少有一个独特的表位,可被狼疮自身抗体识别。