UDP-glucose dehydrogenase from bovine liver: primary structure and relationship to other dehydrogenases.
The primary structure of bovine liver UDP-glucose dehydrogenase (UDPGDH), a hexameric, NAD(+)-linked enzyme, has been determined at the protein level. The 52-kDa subunits are composed of 468 amino acid residues, with a free N-terminus and a Ser/Asn microhetergeneity at one position. The sequence shares 29.6% positional identity with GDP-mannose dehydrogenase from Pseudomonas, confirming a similarity earlier noted between active site peptides. This degree of similarity is comparable to the 31.1% identity vs. the UDPGDH from type A Streptococcus. Database searching also revealed similarities to a hypothetical sequence from Salmonella typhimurium and to "UDP-N-acetyl-mannosaminuronic acid dehydrogenase" from Escherichia coli. Pairwise identities between bovine UDPGDH and each of these sequences were all in the range of approximately 26-34%. Multiple alignment of all 5 sequences indicates common ancestry for these 4-electron-transferring enzymes. There are 27 strictly conserved residues, including a cysteine residue at position 275, earlier identified by chemical modification as the expected catalytic residue of the second half-reaction (conversion of UDP-aldehydoglucose to UDP-glucuronic acid), and 2 lysine residues, at positions 219 and 338, one of which may be the expected catalytic residue for the first half-reaction (conversion of UDP-glucose to UDP-aldehydoglucose). A GXGXXG pattern characteristic of the coenzyme-binding fold is found at positions 11-16, close to the N-terminus as with "short-chain" alcohol dehydrogenases.(ABSTRACT TRUNCATED AT 250 WORDS)
Hempel
J
J
Department of Molecular Genetics and Biochemistry, University of Pittsburgh, Pennsylvania 15261.
Perozich
J
J
Romovacek
H
H
Hinich
A
A
Kuo
I
I
Feingold
D S
DS
D000595:Amino Acid Sequence; D000818:Animals; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry; D002417:Cattle; D004926:Escherichia coli / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000737:chemistry*; D011549:Pseudomonas / Q000201:enzymology; D012486:Salmonella typhimurium / Q000201:enzymology; D017385:Sequence Homology; D013297:Streptococcus pyogenes / Q000201:enzymology; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000737:chemistry*
D010088:Oxidoreductases; D002237:Carbohydrate Dehydrogenases; C023228:UDP-N-acetylmannosamine dehydrogenase; C020562:GDPmannose dehydrogenase; D014533:Uridine Diphosphate Glucose Dehydrogenase