title: Loop interactions during catalysis by dihydrofolate reductase from Moritella profunda.
abstract: Dihydrofolate reductase (DHFR) is often used as a model system to study the relation between protein dynamics and catalysis. We have studied a number of variants of the cold-adapted DHFR from Moritella profunda (MpDHFR), in which the catalytically important M20 and FG loops have been altered, and present a comparison with the corresponding variants of the well-studied DHFR from Escherichia coli (EcDHFR). Mutations in the M20 loop do not affect the actual chemical step of transfer of hydride from reduced nicotinamide adenine dinucleotide phosphate to the substrate 7,8-dihydrofolate in the catalytic cycle in either enzyme; they affect the steady state turnover rate in EcDHFR but not in MpDHFR. Mutations in the FG loop also have different effects on catalysis by the two DHFRs. Despite the two enzymes most likely sharing a common catalytic cycle at pH 7, motions of these loops, known to be important for progression through the catalytic cycle in EcDHFR, appear not to play a significant role in MpDHFR.
authors.affiliation: School of Chemistry, Cardiff University , Park Place, Cardiff CF10 3AT, United Kingdom.
authors.forename: Enas M
authors.initials: EM
authors.lastname: Behiry
authors.forename: Rhiannon M
authors.initials: RM
authors.lastname: Evans
authors.forename: Jiannan
authors.initials: J
authors.lastname: Guo
authors.forename: E Joel
authors.initials: EJ
authors.lastname: Loveridge
authors.forename: Rudolf K
authors.initials: RK
authors.lastname: Allemann
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D055162:Biocatalysis; D004926:Escherichia coli / Q000201:enzymology; D005492:Folic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D034221:Moritella / Q000201:enzymology*; D016297:Mutagenesis, Site-Directed; D009249:NADP / Q000737:chemistry; D010084:Oxidation-Reduction; D011487:Protein Conformation; D013045:Species Specificity; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C010920:dihydrofolate; D009249:NADP; D005492:Folic Acid; D013762:Tetrahydrofolate Dehydrogenase