Pressure denaturation of phosphorylating glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus.
The effects of hydrostatic pressure on apo wild-type glyceraldehyde-3-phosphate dehydrogenase (wtGAPDH) from Bacillus stearothermophilus (B. stearothermophilus) have been studied by fluorescence spectroscopy under pressure from 0.1 to 650 MPa. Unlike yeast GAPDH [Ruan, K. C., and Weber, G. (1989) Biochemistry 28, 2144-2153], denaturation of the tetrameric apo wtGAPDH from B. stearothermophilus is likely to precede dissociation into subunits. As expected, denaturation is accompanied by the loss of enzymatic activity. B. stearothermophilus apo wtGAPDH interfaces are less pressure sensitive than apo yeast GAPDH ones, while NAD does not protect B. stearothermophilus wtGAPDH against denaturation by pressure. The pressure effects on B. stearothermophilus GAPDH whose R and Q-axis interfaces were destabilized by disruption of interfacial hydrogen bonds are similar to that of apo wtGAPDH.
Roitel
O
O
Maturation des ARN et Enzymologie Moléculaire, UMR 7567 CNRS-Université Henri Poincaré, Nancy I, Boulevard des Aiguillettes, Vandoeuvre-lès-Nancy, 54506, France.
Bec
N
N
Lange
R
R
Balny
C
C
Branlant
G
G
D001051:Apoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006860:Hydrogen Bonding; D006874:Hydrostatic Pressure; D009154:Mutation; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence
D001051:Apoenzymes; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases