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NMR‐derived folate‐bound structure of dihydrofolate reductase 1 from the halophile Haloferax volcanii
Author(s) -
Boroujerdi Arezue F. B.,
Young John K.
Publication year - 2009
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.21096
Subject(s) - dihydrofolate reductase , chemistry , haloferax volcanii , halophile , nuclear magnetic resonance spectroscopy , two dimensional nuclear magnetic resonance spectroscopy , stereochemistry , crystallography , resonance (particle physics) , enzyme , biochemistry , physics , biology , genetics , bacteria , archaea , gene , particle physics
Abstract Folate binds to dihydrofolate reductase (DHFR) to form a binary complex whose structure maintains the overall configuration of the enzyme; however, some significant changes are evident when a comparison is made to the enzyme. The structure of DHFR1 from the halophilic Halopherax volcanii was solved in its folate‐bound form using nuclear magnetic resonance spectroscopy. NOE data obtained from the 15 N‐NOESY‐HSQC and 13 C‐NOESY‐HSQC experiments of the triply labeled ( 1 H, 13 C, and 15 N) binary complex were used as input for the structure calculation with the Crystallography and Nuclear Magnetic Resonance System program. The resulting family of structures was compared with the enzyme solved by both nuclear magnetic resonance and X‐ray crystallography and also to the mesophilic folate‐bound enzyme from Escherichia coli . The binary complex exhibited less convergence of structure in the α2‐helix and differences in the hinge residues D39 and A94. In comparison to the previously reported mesophilic binary complex solved by X‐ray crystallography, the halophilic binary complex reported here does not agree with the convergence of the M20 loop to a single structure. The corresponding L21 loop of the halophilic binary complex family of structures solved by nuclear magnetic resonance indicates variability in this region. © 2008 Wiley Periodicals, Inc. Biopolymers 91: 140–144, 2009. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com