Beyond the catalytic core of ALDH: a web of important residues begins to emerge.
Site-directed mutagenesis was performed in class 3 aldehyde dehydrogenase (ALDH) on both strictly conserved, non-glycine residues, Glu-333 and Phe-335. Both lie in Motif 8 and are indicated to be of central catalytic importance from their positions in the tertiary structure. In addition, a highly conserved residue at the end of Motif 8, Pro-337, and Asp-247, which interacts with the main chain of Motif 8, were also mutated. All substitutions were conservative. Kinetic values clearly show that Glu-333 and Phe-335 are crucial to efficient catalysis, along with Asp-247. Pro-337 appears to have a different role, most likely relating to folding.
Hempel
J
J
Department of Biological Sciences, University of Pittsburgh, 301 Clapp Hall, 15260, Pittsburgh, PA, USA. hempel@psc.edu
Lindahl
R
R
Perozich
J
J
Wang
B
B
Kuo
I
I
Nicholas
H
H
D000444:Aldehyde Dehydrogenase / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs; D000818:Animals; D020134:Catalytic Domain; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D017510:Protein Folding; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D000444:Aldehyde Dehydrogenase