Cloning and sequence analysis of a cDNA for 3-hydroxyisobutyrate dehydrogenase. Evidence for its evolutionary relationship to other pyridine nucleotide-dependent dehydrogenases.
A 1.7-kilobase pair cDNA clone encoding 3-hydroxyisobutyrate dehydrogenase has been isolated by screening a rat liver lambda gt11 library with a 17-base oligonucleotide probe which corresponds to a portion of the N-terminal amino acid sequence of rabbit liver 3-hydroxyisobutyrate dehydrogenase. The cDNA contains an open reading frame of 1038 base pairs which includes an amino acid sequence that matches the N-terminal 35 amino acid sequence of rabbit 3-hydroxyisobutyrate dehydrogenase at 33 residues. The cDNA predicts a 300-amino acid mature protein with an amino acid composition and molecular weight very similar to that of rabbit liver 3-hydroxyisobutyrate dehydrogenase. Northern blot analysis of total RNA from several rat tissues shows an mRNA of approximately 2.0 kilobase pairs in each tissue. Relative mRNA levels were: kidney greater than liver = heart greater than muscle. The amino acid sequence of 3-hydroxyisobutyrate dehydrogenase shows similarity to several other pyridine nucleotide-dependent dehydrogenases. The resemblance to malate and lactate dehydrogenases suggests that the nucleotide-binding domain is located in the N-terminal region of the protein.
Rougraff
P M
PM
Department of Biochemistry, Indiana University School of Medicine, Indianapolis 46223.
Zhang
B
B
Kuntz
M J
MJ
Harris
R A
RA
Crabb
D W
DW
D000429:Alcohol Oxidoreductases / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005796:Genes; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010088:Oxidoreductases / Q000235:genetics*; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D051381:Rats; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D012333:RNA, Messenger; D004247:DNA; D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; C021398:3-hydroxyisobutyrate dehydrogenase