15N and 1H NMR evidence for multiple conformations of the complex of dihydrofolate reductase with its substrate, folate

FEBS Letters

Crase, J.¹  De Graw, J.¹  Roberts, G.C.K.⁵  Colwell, W.T.¹  Birdsall, B.³  Feeney, J.³  Hammond, S.⁴  Searle, M.S.² 

[1] Bio-Organic Chemistry Laboratory, SRI International, Menlo Park, CA 94025, USA;Cancer Institute, 481 Little Lonsdale Street, Melbourne, Victoria 3000, Australia;Division of Physical Biochemistry, National Institute for Medical Research, Mill Hill, London NW7 1AA, England;Analytical Chemistry Division, Shell Research Ltd, Sittingbourne Research Centre, Sittingbourne ME9 8AG, England;Department of Biochemistry, University of Leicester, Leicester LE1 7RH, England

关键词: Dihydrofolate reductase;    Folate;    15N-NMR;    2D NMR;   

DOI  :  10.1016/0014-5793(87)81252-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The binding of folate to Lactobacillus casei dihydrofolate reductase in the presence and absence of NADP⁺ has been studied by ¹⁵N NMR, using [5-¹⁵N]folate. In the presence of NADP⁺, three separate signals were observed for the single ¹⁵N atom, in agreement with our earlier evidence from ¹H and ¹³C NMR for multiple conformations of this complex [(1982) Biochemistry 21, 5831–5838]. The ¹⁵N spectra of the binary enzymefolate complex provide evidence for the first time that this complex also exists in at least two conformational states. This is confirmed by the observation of two separate resonances for the 7-proton of bound folate, located by two-dimensional exchange spectroscopy.

【 授权许可】

Unknown