Engineering thermostability in archaebacterial glyceraldehyde-3-phosphate dehydrogenase. Hints for the important role of interdomain contacts in stabilizing protein conformation.
Construction of hybrid enzymes between the glyceraldehyde-3-phosphate dehydrogenases from the mesophilic Methanobacterium bryantii and the thermophilic Methanothermus fervidus by recombinant DNA techniques revealed that a short C-terminal fragment of the Mt. fervidus enzyme contributes largely to its thermostability. This C-terminal region appears to be homologous to the alpha 3-helix of eubacterial and eukaryotic glyceraldehyde-3-phosphate dehydrogenases which is involved in the contacts between the two domains of the enzyme subunit. Site-directed mutagenesis experiments indicate that hydrophobic interactions play an important role in these contacts.
Biro
J
J
Max-Planck-Institut für Biochemie, Martinsried, FRG.
Fabry
S
S
Dietmaier
W
W
Bogedain
C
C
Hensel
R
R
D000595:Amino Acid Sequence; D001105:Archaea / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D005818:Genetic Engineering; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D006358:Hot Temperature; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D011489:Protein Denaturation; D013329:Structure-Activity Relationship
D009841:Oligonucleotides; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases