Probing antibody diversity by 2D NMR: comparison of amino acid sequences, predicted structures, and observed antibody-antigen interactions in complexes of two antipeptide antibodies.
Probing antibody diversity by 2D NMR: comparison of amino acid sequences, predicted structures, and observed antibody-antigen interactions in complexes of two antipeptide antibodies.
复制标题
通过 2D NMR 探测抗体多样性：比较两种抗肽抗体复合物中的氨基酸序列、预测结构和观察到的抗体-抗原相互作用。
作者：
R. Levy;O. Assulin;T. Scherf;M. Levitt;J. Anglister
两种单克隆抗体（TE32 和 TE33）的芳香族氨基酸与霍乱毒素肽 (CTP3) 的特定氨基酸残基之间的相互作用已通过二维 (2D) 转移 NOE 差异光谱法确定。发现芳香族氨基酸在肽结合中发挥重要作用。在两种抗体中，两个色氨酸、两个酪氨酸残基和一个组氨酸残基与肽相互作用。 TE33 中还有一个额外的苯丙氨酸残基也与肽相互作用。已发现与抗体相互作用的 CTP3 肽残基是 val 3、pro 4、gly 5、gln 7、his 8 和 asp 10。我们通过直接 mRNA 测序确定了两种抗体的氨基酸序列。计算机分子模型已用于根据其他抗体的已知构象构建这两种抗体的详细全原子模型。这些模型允许明确分配与肽相互作用的大多数抗体残基。将两种抗CTP3抗体与来自同一基因家族的其他抗体的氨基酸序列进行比较表明，尽管这些抗体具有不同的特异性，但参与CTP3结合的大多数芳香族残基是保守的。这种相似性表明这些芳香族残基形成了一个通用的疏水口袋，而互补决定区（CDR）中的其他残基则调节结合位点的形状和极性以适应特定的抗原。
The interactions between the aromatic amino acids of two monoclonal antibodies (TE32 and TE33) with specific amino acid residues of a peptide of cholera toxin (CTP3) have been determined by two-dimensional (2D) transferred NOE difference spectroscopy. Aromatic amino acids are found to play an important role in peptide binding. In both antibodies two tryptophan and two tyrosine residues and one histidine residue interact with the peptide. In TE33 there is an additional phenylalanine residue that also interacts with the peptide. The residues of the CTP3 peptide that have been found to interact with the antibody are val 3, pro 4, gly 5, gln 7, his 8, and asp 10. We have determined the amino acid sequences of the two antibodies by direct mRNA sequencing. Computerized molecular modeling has been used to build detailed all-atom models of both antibodies from the known conformations of other antibodies. These models allow unambiguous assignment of most of the antibody residues that interact with the peptide. A comparison of the amino acid sequences of the two anti-CTP3 antibodies with other antibodies from the same gene family reveals that the majority of the aromatic residues involved in the binding of CTP3 are conserved although these antibodies have different specificities. This similarity suggests that these aromatic residues create a general hydrophobic pocket and that other residues in the complementarity-determining regions (CDRs) modulate the shape and the polarity of the combining site to fit the specific antigens.
色氨酸残基对单克隆抗二硝基苯基自旋标记抗体结合位点的贡献。
影响因子：
2.9
作者：
Anglister,J;Bond,MW;Frey,T;Leahy,D;Levitt,M;McConnell,HM;Rule,GS;Tomasello,J;Whittaker,M
通讯作者：
Whittaker,M