STEREOSPECIFIC ASSIGNMENTS OF THE LEUCINE METHYL RESONANCES IN THE H-1-NMR SPECTRUM OF LACTOBACILLUS-CASEI DIHYDROFOLATE-REDUCTASE

被引：28

作者：

OSTLER, G

SOTERIOU, A

MOODY, CM

KHAN, JA

BIRDSALL, B

CARR, MD

YOUNG, DW

FEENEY, J

机构：

[1] NATL INST MED RES,MOLEC STRUCT LAB,MILL HILL,LONDON NW7 1AA,ENGLAND

[2] UNIV SUSSEX,SCH CHEM & MOLEC SCI,BRIGHTON BN1 9QJ,E SUSSEX,ENGLAND

来源：

FEBS LETTERS | 1993年 / 318卷 / 02期

关键词：

DIHYDROFOLATE REDUCTASE; STEREOSPECIFIC NMR ASSIGNMENT; DEUTERATED LEUCINE;

D O I：

10.1016/0014-5793(93)80016-N

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：

A general method is described for the stereospecific assignment of methyl resonances in protein NMR spectra based on selective deuteration procedures. A selectively deuterated dihydrofolate reductase from L. casei was prepared by incorporating stereoselectively deuterated L-leucine, (2S,4R)[5,5,5-H-2(3)]leucine. By comparing the COSY spectra of the dihydrofolate reductase-methotrexate complexes formed using deuterated and non-deuterated enzyme the stereospecific assignments for resonances of all 13 leucine residues were obtained by noting the absence of cross-peaks in spectra from the deuterated proteins.

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页码：177 / 180

页数：4

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