Occurrence of beta-alanine-specific opine dehydrogenase in the muscle of the limpet Cellana grata Gould (Archaeogastropoda).
The muscular tissues of the limpet Cellana grata exhibited beta-alanopine dehydrogenase (beta-AlDH) activity in addition to tauropine dehydrogenase (TaDH) activity and weak lactate dehydrogenase activity. Opine dehydrogenases (OpDHs) were purified, and two different types of OpDH, i.e. TaDHs and OpDHs showing beta-AlDH activity, were isolated. From the specificity for amino acid and opine, OpDHs showing beta-AlDH activity were concluded to be a true beta-AlDH showing strict substrate specificity for beta-alanine. Although the catalytic properties of beta-AlDH and TaDH were essentially similar, they were distinct from each other with respect to the amino acid substrate specificity and the K(m) values. Apparent K(m) values (mM) for the preferred amino acid substrate, pyruvate, NADH, the preferred opine substrate, and NAD+ were: 14.3 (beta-alanine), 0.19, 0.032, 35.2 (beta-alanopine), and 0.78 for beta-AlDH; and 33.3 (taurine), 0.53, 0.076, 48.6 (tauropine), and 0.58 for TaDH, respectively. Great similarities were found between beta-AlDH and TaDH with respect to molecular properties: molecular masses (both enzymes were monomeric proteins of approximately 40,000 Da), amino acid compositions, and N-terminal amino acid sequences (30 amino acid residues were identical). Partial similarities were also recognized between their lysyl endopeptidase maps. These results clearly show that beta-alanine-specific OpDH, a true beta-AlDH, is present in the limpet muscle.
Kan-no
N
N
Department of Marine Biochemistry, School of Fisheries Sciences, Kitasato University, Iwate, Japan. kan-no@nnet.ne.jp
Sato
M
M
Yokoyama
T
T
Nagahisa
E
E
D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008974:Mollusca / Q000201:enzymology*; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010449:Peptide Mapping; D017421:Sequence Analysis; D012697:Serine Endopeptidases; D013379:Substrate Specificity; D015091:beta-Alanine / Q000378:metabolism*
D000596:Amino Acids; D015091:beta-Alanine; D000594:Amino Acid Oxidoreductases; D000587:Oxidoreductases Acting on CH-NH Group Donors; C030014:alanopine dehydrogenase; C060511:opine dehydrogenase, NAD+; C050564:tauropine dehydrogenase; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase