Optimising selective deuteration of proteins for 2D 1H NMR detection and assignment studies Application to the Phe residues of Lactobacillus casei dihydrofolate reductase

FEBS Letters

Ostler, G.²  Arnold, J.R.P.¹  Roberts, G.C.K.¹  Tendler, S.J.B.²  Birdsall, B.²  Akiboye, J.²  Feeney, J.²  Barbero, J.Jiménez²  Kühn, A.³  Roth, K.³ 

[1] Biochemistry Department, University of Leicester University Road, Leicester LE1 7RH, England;Physical Biochemistry Division, NIMR, Mill Hill, London NW7 1AA, England;Institut für Organische Chemie, Freie Universität, Berlin WE 02, Germany

关键词: NMR;    2D;    Dihydrofolate reductase;    Selective deuteration;   

DOI  :  10.1016/0014-5793(89)80431-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A selectively deuterated dihydrofolate reductase from L. casei has been prepared containing partially deuterated aromatic amino acids. This provides simplified 2D NMR spectra and allows signals from all 8 Phe residues to be identified. The pattern of deuteration is such that (i) the only cross-peaks detected in the aromatic region of the 2D COSY spectrum are those between the Phe 2′,6′ and 3′,5′ protons and (ii) chemical shift degeneracy in the aromatic region is removed thus allowing unambiguous assignment of cross-peaks in 2D NOESY spectra required for specific assignment purposes.

【 授权许可】

Unknown