title: High pressure nmr study of dihydrofolate reductase from a deep-sea bacterium Moritella profunda.
abstract: We have investigated the effect of pressure and temperature on the structural and thermodynamic stability of a protein dihydrofolate reductase from a deep-sea bacterium Moritella profunda in its folate-bound form in the pressure range between 3 and 375 MPa and the temperature range between -5 and 30 degrees C. The on-line cell variable pressure 1H NMR spectroscopy has been used to analyze the chemical shift and signal intensity in one-dimensional 1H NMR spectra. Thermodynamic analysis based on signal intensities from protons in the core part indicates that the thermodynamic stability of Moritella profunda DHFR is relatively low over the temperature range between -5 and 30 degrees C (deltaG0=15.8 +/- 4.1 kJ/mol at 15 degrees C), but is well adapted to the living environment of the bacterium (2 degrees C and 28 MPa), with the maximum stability around 5 degrees C (at 0.1 MPa) and a relatively small volume change upon unfolding (deltaV= 66 +/- 19 ml/mol). Despite the relatively low overall stability, the conformation in the core part of the folded protein remains intact up to approximately 200 MPa, showing marked stability of the core of this protein.
authors.affiliation: Department of Biotechnological Science, School of Biology-Oriented Science and Technology, Kinki University, Wakayama, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Hata
authors.forename: R
authors.initials: R
authors.lastname: Kono
authors.forename: M
authors.initials: M
authors.lastname: Fujisawa
authors.forename: R
authors.initials: R
authors.lastname: Kitahara
authors.forename: Y O
authors.initials: YO
authors.lastname: Kamatari
authors.forename: K
authors.initials: K
authors.lastname: Akasaka
authors.forename: Y
authors.initials: Y
authors.lastname: Xu
mesh_terms: D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000737:chemistry; D004795:Enzyme Stability; D009682:Magnetic Resonance Spectroscopy; D008386:Marine Biology; D034221:Moritella / Q000201:enzymology* / Q000502:physiology; D011312:Pressure; D013696:Temperature; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry*; D013816:Thermodynamics
chemical_list: D001426:Bacterial Proteins; D013762:Tetrahydrofolate Dehydrogenase