IDENTIFICATION AND CHEMICAL SYNTHESIS OF A RIBOSOMAL-PROTEIN ANTIGENIC DETERMINANT IN SYSTEMIC LUPUS-ERYTHEMATOSUS
IDENTIFICATION AND CHEMICAL SYNTHESIS OF A RIBOSOMAL-PROTEIN ANTIGENIC DETERMINANT IN SYSTEMIC LUPUS-ERYTHEMATOSUS
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DOI：
10.1073/pnas.83.19.7419
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发表时间：
1986-10-01
影响因子：
11.1
通讯作者：
BROT, N
中科院分区：
文献类型：
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作者：
ELKON, K;SKELLY, S;BROT, N
用等电聚焦和阴离子交换层析法分离了系统性红斑狼疮（SLE）患者血清中的真核核糖体蛋白P0、P1和P2（P蛋白）及其抗原决定簇。经SephadexG-200柱层析测定，复合物的表观分子量为140 kDa。因此，P0可能是大肠杆菌核糖体蛋白L10的真核等价物。此外，在HeLa细胞的postribosomal上清液中检测到所有三种P蛋白，并且发现P0和P1比它们的核糖体结合的对应物更酸性。部分蛋白水解实验表明，SLE抗P血清识别的P2等效蛋白从卤虫（eL 12）的一端或两端。含有P0、P1和P2抗体的16份SLE血清与eL 12的羧基末端22个氨基酸的肽反应，而不与20个氨基酸的氨基末端肽反应。尽管羧基末端肽完全抑制了抗血清在免疫印迹上与所有三种蛋白质反应的能力，但同一肽仅使SLE抗体与天然非变性P蛋白的结合略有减少。这些发现表明，SLE抗P抗体与所有三种P蛋白上的单个连续（线性）抗原决定簇反应，但其他抗体识别构象决定簇。
The characteristics of eukaryotic ribosomal proteins P0, P1, and P2 (P proteins) and their antigenic determinants were studied using the sera of patients with systemic lupus erythematosus (SLE) P0, P1, and P2 were isolated as a macromolecular complex by preparative isoelectric focusing and anion-exchange chromatography in the presence of 6 M urea. The apparent molecular size of the complex was 140 kDa as determined by gel filtration on Sephadex G-200 column. P0 may, therefore, be the eukaryotic equivalent of Escherichia coli ribosomal protein L10. In addition, all three P proteins were detected in the postribosomal supernatant of HeLa cells, and P0 and P1 were found to be more acidic than their ribosome-bound counterparts. Partial proteolysis experiments revealed that SLE anti-P sera recognized one or both ends of the P2 equivalent protein from Artemia salina (eL12). Sixteen SLE sera containing antibodies to P0, P1, and P2 reacted with a carboxyl-terminal peptide 22 amino acids in length of eL12 and not with an amino-terminal peptide of 20 amino acids. Even though the carboxyl-terminal peptide completely inhibited the ability ofthe antiserum to react with all three proteins on an immunological blot the same peptide produced only small decreases in binding of the SLE antibody to the native, nondenatured P proteins. These findings indicate that SLE anti-P antibodies react with a single sequential (linear) antigenic determinant on all three P proteins, but that additional antibodies recognize a conformational determinant(s).