与亲环蛋白结合的环孢菌素A的同位素编辑核磁共振：反式9,10酰胺键的证据。

Isotope-edited NMR of cyclosporin A bound to cyclophilin: evidence for a trans 9,10 amide bond.

作者信息

Fesik S W, Gampe R T, Holzman T F, Egan D A, Edalji R, Luly J R, Simmer R, Helfrich R, Kishore V, Rich D H

机构信息

Pharmaceutical Discovery Division, Abbott Laboratories, Abbott Park, IL 60064.

出版信息

Science. 1990 Dec 7;250(4986):1406-9. doi: 10.1126/science.2255910.

DOI:10.1126/science.2255910

PMID:2255910

Abstract

The binding of a 13C-labeled cyclosporin A (CsA) analog to cyclophilin (peptidyl prolyl isomerase) was examined by means of isotope-edited nuclear magnetic resonance (NMR) techniques. A trans 9,10 peptide bond was adopted when CsA was bound to cyclophilin, in contrast to the cis 9,10 peptide bond found in the crystalline and solution conformations of CsA. Furthermore, nuclear Overhauser effects (NOEs) were observed between the zeta 3 and epsilon 3 protons of the methylleucine (MeLeu) residue at position 9 of CsA and tryptophan121 (Trp121) and phenylalanine (Phe) protons of cyclophilin, suggesting that the MeLeu9 residue of CsA interacts with cyclophilin. These results illustrate the power of isotope-edited NMR techniques for rapidly providing useful information about the conformations and active site environment of inhibitors bound to their target enzymes.

摘要

通过同位素编辑核磁共振（NMR）技术检测了13C标记的环孢菌素A（CsA）类似物与亲环蛋白（肽基脯氨酰异构酶）的结合。当CsA与亲环蛋白结合时，采用反式9,10肽键，这与CsA晶体和溶液构象中发现的顺式9,10肽键相反。此外，在CsA第9位甲基亮氨酸（MeLeu）残基的ζ3和ε3质子与亲环蛋白的色氨酸121（Trp121）和苯丙氨酸（Phe）质子之间观察到核Overhauser效应（NOE），表明CsA的MeLeu9残基与亲环蛋白相互作用。这些结果说明了同位素编辑NMR技术在快速提供有关与其靶酶结合的抑制剂的构象和活性位点环境的有用信息方面的作用。