Naturally processed peptides from rheumatoid arthritis associated and non-associated HLA-DR alleles.
Naturally processed peptides from rheumatoid arthritis associated and non-associated HLA-DR alleles.
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DOI：
10.4049/jimmunol.155.12.5655
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发表时间：
1995-12
影响因子：
4.4
通讯作者：
D. Kirschmann;K. Duffin;C. E. Smith;J. K. Welply;S. Howard;B. Schwartz;S. Woulfe
中科院分区：
文献类型：
--
作者：
D. Kirschmann;K. Duffin;C. E. Smith;J. K. Welply;S. Howard;B. Schwartz;S. Woulfe
通过毛细管液相色谱和质谱分析来自免疫亲和纯化的HLA-DRB 1 *0401、-DRB 1 *0404（类风湿性关节炎（RA）相关）和-DRB 1 *0402（非RA相关）分子的天然加工肽。从每种HLA-DR蛋白质中观察到的60多种洗脱肽的分子量范围为788至3535原子质量单位，对应于长度为7至32个氨基酸的肽。对60多个丰富的肽进行测序，揭示了仅来自12种不同蛋白质的嵌套肽组。这些蛋白质中的大多数是膜相关的（HLA I类、II类和IG分子）。对应于内源性肽序列的合成肽以高亲和力（5至80 nM）结合至它们从中洗脱的HLA-DR分子。此外，大多数是混杂的结合肽，因为它们也与其他HLA-DR分子结合。洗脱肽序列的截短和麻风分枝杆菌肽的丙氨酸扫描突变分析用于鉴定参与与DRB 1 *0404和DRB 1 *0402分子结合的肽残基。此外，从DRB 1 *0402分子中洗脱出不变链肽，但不从RA相关分子中洗脱。DRB 1 *0401和DRB 1 *0404分子中缺乏恒定链肽可能有助于将自身抗原肽加载到这些分子中并与疾病相关。
Naturally processed peptides from immunoaffinity-purified HLA-DRB1*0401, -DRB1*0404 (rheumatoid arthritis (RA)-associated), and -DRB1*0402 (non-RA-associated) molecules were analyzed by capillary liquid chromatography and mass spectrometry. The molecular weights observed for more than 60 eluted peptides from each HLA-DR protein ranged from 788 to 3535 atomic mass units, corresponding to peptides 7 to 32 amino acids in length. Sequencing of more than 60 of the abundant peptides revealed nested sets of peptides that were derived from only 12 different proteins. The majority of these proteins were membrane-associated (HLA class I, class II, and Ig molecules). Synthetic peptides, corresponding to endogenous peptide sequences, bound with high affinity (5 to 80 nM) to the HLA-DR molecules from which they were eluted. In addition, most were promiscuous binding peptides in that they also bound to other HLA-DR molecules. Truncations of eluted peptide sequences and alanine scanning mutational analysis of a Mycobacterium leprae peptide were used to identify the peptide residues involved in binding to DRB1*0404 and DRB1*0402 molecules. Furthermore, an invariant chain peptide was eluted from the DRB1*0402 molecules but not from the RA-associated molecules. The lack of invariant chain peptides from DRB1*0401 and DRB1*0404 molecules may contribute to the loading of autoantigen peptides into these molecules and to their association with disease.