Definition of specific peptide motifs for four major HLA-A alleles.
Definition of specific peptide motifs for four major HLA-A alleles.
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DOI：
10.4049/jimmunol.152.8.3913
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发表时间：
1994-04
影响因子：
4.4
通讯作者：
Ralph T. Kubo;Alessandro Sette;Howard M. Grey;Ettore Appella;Kazuyasu Sakaguchi;N. Zhu;D. Arnott;Nicholas E. Sherman;J. Shabanowitz;Hanspeter Michel
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文献类型：
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作者：
Ralph T. Kubo;Alessandro Sette;Howard M. Grey;Ettore Appella;Kazuyasu Sakaguchi;N. Zhu;D. Arnott;Nicholas E. Sherman;J. Shabanowitz;Hanspeter Michel
用三种互补的方法鉴定了人类MHC I类分子的等位基因特异性基序，即人类MHC-A1、A3、A11和A24。首先，对亲和纯化的I类分子的酸洗脱肽库进行氨基酸序列分析，确定了9个或10个氨基酸的推测基序，并在第2位和COOH末端含有关键的锚定残基。这些基序是不同的，除了人类白细胞抗原-A3和A11基序彼此非常相似。其次，通过分析聚丙氨酸类似物与纯化的人类白细胞抗原-A分子的结合能力，验证了这些假定基序的正确性。确定了几个可选的锚基残基，这些残基在汇集的多肽序列分析中并不明显。第三，用串联质谱法测定了从人类白细胞抗原A1、A11和A24洗脱的单肽的序列。虽然也鉴定出了长度为8、10、11和12个氨基酸的多肽，但仍以九聚体为主。这些多肽显示由第2位和COOH末端的特定基序预测的锚定残基，与多肽长度无关。自然处理的多肽的合成版本被证明与适当的人类白细胞抗原-A等位基因结合，IC50值在0.3-200-NM范围内。一种合理的方法来搜索具有已知氨基酸序列的AGS，以寻找受一些最常见的HLA-A类型限制的表位，并且具有潜在的临床重要性。
Allele-specific motifs for the human MHC class I molecules, HLA-A1, A3, A11, and A24 were characterized by three complementary approaches. First, amino acid sequence analysis of acid eluted peptide pools from affinity purified class I molecules defined putative motifs 9 or 10 amino acids in length and bearing critical anchor residues at position 2 and at the COOH-terminal. These motifs were distinct, with the exception of the HLA-A3 and A11 motifs that were very similar to each other. Second, the correctness of these putative motifs was verified by analyzing the binding capacity of polyalanine peptide analogues to purified HLA-A molecules. Several alternative anchor residues that were not obvious from the pooled peptide sequencing analysis were identified. Third, sequences of individual peptides eluted from HLA-A1, A11, and A24 were determined by tandem mass spectrometry. Nonamers were the predominant species, although peptides of 8, 10, 11, and 12 amino acids in length were also identified. These peptides displayed anchor residues predicted by the specific motifs at position 2 and at the COOH-terminal, regardless of peptide length. Synthetic versions of the naturally processed peptides were shown to bind to the appropriate HLA-A alleles with IC50 values in the 0.3- to 200-nM range. A rational approach to search Ags with known amino acid sequences for epitopes restricted by some of the most common HLA-A types and of potential clinical importance is now feasible.