15N NMR studies of the conformation ofE. colidihydrofolate reductase in complex with folate or methotrexate
- 9 September 1991
- journal article
- Published by Wiley in FEBS Letters
- Vol. 289 (2) , 231-234
- https://doi.org/10.1016/0014-5793(91)81077-l
Abstract
We have employed 15N NMR to characterize the conformations of Escherichia coli dihydrofolate reductase (ECDHFR) in complex with [5-15N]folate or [5-15N]methotrexate (MTX). Two 15N resonances were observed for DHFR/MTX binary complex. The relative population of these two conformations is pH dependent. Addition of NADP+ or NADPH results in the disappearance of the low field resonance. In contrast, only one conformation was observed for both the DHFR/folate and DHFR/folate/NADP+ complexes. However, the 15N chemical shift of [5-15N]folate in the binary DHFR/folate complex is 7.28 ppm upfield from that of the ternary complex, suggesting the possible loss of a hydrogen bonding to N5 of folate in the ternary complexKeywords
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