SEQUENCE-SPECIFIC H-1-NMR ASSIGNMENTS FOR THE AROMATIC REGION OF SEVERAL BIOLOGICALLY-ACTIVE, MONOMERIC INSULINS INCLUDING NATIVE HUMAN INSULIN

被引:19
作者
ROY, M
LEE, RW
KAARSHOLM, NC
THOGERSEN, H
BRANGE, J
DUNN, MF
机构
[1] UNIV CALIF RIVERSIDE,DEPT BIOCHEM,RIVERSIDE,CA 92521
[2] UNIV CALIF RIVERSIDE,DEPT CHEM,RIVERSIDE,CA 92521
[3] NOVO RES INST,DK-2880 BAGSVAERD,DENMARK
关键词
(Human); Insulin; Monomeric insulin mutant; NMR; !sup]1[!/sup]H-; Sequence-specific assignment; Two-dimensional COSY;
D O I
10.1016/0167-4889(90)90027-B
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The aromatic region of the 1H-FT-NMR spectrum of the biologically fully-potent, monomeric human insulin mutant, B9 Ser → Asp, B27 Thr → Glu has been investigated in D2O. At 1 to 5 mM concentrations, this mutant insulin is monomeric above pH 7.5. Coupling and amino acid classification of all aromatic signals is established via a combination of homonuclear one- and two-dimensional methods, including COSY, multiple quantum filters, selective spin decoupling and pH titrations. By comparisons with other insulin mutants and with chemically modified native insulins, all resonances in the aromatic region are given sequence-specific assignments without any reliance on the various crystal structures reported for insulin. These comparisons also give the sequence-specific assignments of most of the aromatic resonances of the mutant insulins B16 Tyr → Glu, B27 Thr → Glu and B25 Phe → Asp and the chemically modified species des-(B23-B30) insulin and monoiodo-Tyr A14 insulin. Chemical dispersion of the assigned resonances, ring current perturbations and comparisons at high pH have made possible the assignment of the aromatic resonances of human insulin, and these studies indicate that the major structural features of the human insulin monomer (including those critical to biological function) are also present in the monomeric mutant. © 1990.
引用
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页码:63 / 73
页数:11
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