NMR study of the complexes between a synthetic peptide derived from the B subunit of cholera toxin and three monoclonal antibodies against it.
NMR study of the complexes between a synthetic peptide derived from the B subunit of cholera toxin and three monoclonal antibodies against it.
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对源自霍乱毒素 B 亚基的合成肽与三种针对该毒素的单克隆抗体之间的复合物进行 NMR 研究。
DOI：
10.1021/bi00402a034
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
R. Arnon
中科院分区：
文献类型：
--
作者：
J. Anglister;C. Jacob;O. Assulin;G. Ast;R. Pinker;R. Arnon
用核磁共振研究了合成肽与三种不同的抗肽单克隆抗体之间的接触相互作用。该合成肽是CTP 3（霍乱毒素B亚单位的50-64位残基），被认为是抗霍乱的合成疫苗的可能表位。用CTP 3免疫后衍生的杂交瘤细胞系TE 33和TE 32产生与天然毒素交叉反应的抗体。细胞系TE 34产生不结合毒素的抗CTP 3抗体。抗体的选择性氘化已被用于简化质子NMR光谱并将共振分配给特定类型的氨基酸。肽-Fab复合物的质子NMR谱与Fab的质子NMR谱之间的差异谱表明TE 32和TE 33的结合位点结构非常相似，但与TE 34的结合位点结构有很大不同。通过用选择性氘化的抗体Fab片段进行磁化转移实验，我们发现在TE 32和TE 33中，肽的组氨酸残基被埋在抗体结合位点的疏水口袋中，该疏水口袋由色氨酸和两个酪氨酸残基形成。口袋的疏水性质进一步通过对结合的肽组氨酸的C4 H的化学位移缺乏任何pH滴定效应来证明。相反，对于TE 34，我们发现只有一个酪氨酸残基与肽的组氨酸接触。（250字处删节）
The contact interactions between a synthetic peptide and three different anti-peptide monoclonal antibodies have been studied by nuclear magnetic resonance (NMR). The synthetic peptide is CTP3 (residues 50-64 of the B subunit of cholera toxin) suggested as a possible epitope for synthetic vaccine against cholera. The hybridoma cell lines TE33 and TE32 derived after immunization with CTP3 produce antibodies cross-reactive with the native toxin. The cell line TE34 produces anti-CTP3 antibodies that do not bind the toxin. Selective deuteriation of the antibodies has been used to simplify the proton NMR spectra and to assign resonances to specific types of amino acids. The difference spectra between the proton NMR spectrum of the peptide-Fab complex and that of Fab indicate that the combining site structures of TE32 and TE33 are very similar but differ considerably from the combining site structure of TE34. By magnetization transfer experiments with selectively deuteriated Fab fragment of the antibody, we have found that in TE32 and TE33 the histidine residue of the peptide is buried in a hydrophobic pocket of the antibody combining site, formed by a tryptophan and two tyrosine residues. The hydrophobic nature of the pocket is further demonstrated by the lack of any pH titration effect on the chemical shift of the C4H of the bound peptide histidine. In contrast, for TE34 we have found only one tyrosine residue in contact with the histidine of the peptide.(ABSTRACT TRUNCATED AT 250 WORDS)
特定单克隆抗体结合位点中酪氨酸残基与自旋标记半抗原的距离。
影响因子：
2.9
作者：
Anglister,J;Frey,T;McConnell,HM
通讯作者：
McConnell,HM
单克隆抗自旋标记抗体的结合位点区域中的非芳香族氨基酸。
影响因子：
2.9
作者：
Frey,T;Anglister,J;McConnell,HM
通讯作者：
McConnell,HM
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