Structural and Enzymatic Properties of a Gastric NADP(H)- dependent and Retinal-active Alcohol Dehydrogenase

• Published in: The Journal of biological chemistry 1999-09, Vol.274 (37), p.26021-26026
• Main Authors: Peralba, Josep Maria, Cederlund, Ella, Crosas, Bernat, Moreno, Alberto, Julià, Pere, Martínez, Susana Eva, Persson, Bengt, Farrés, Jaume, Parés, Xavier, Jörnvall, Hans
• Format: Article
• Language: English
• Subjects: Alcohol Dehydrogenase - chemistry; Alcohol Dehydrogenase - genetics; Alcohol Dehydrogenase - metabolism; Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; DNA, Complementary; Freshwater; Humans; Kinetics; Medicin och hälsovetenskap; Models, Molecular; Molecular Sequence Data; NADP - metabolism; Protein Conformation; Rana perezi; Ranidae; Retinaldehyde - metabolism; Stomach - enzymology; Substrate Specificity
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.274.37.26021

A class IV-type, gastric alcohol dehydrogenase (ADH) has been purified from frog (Rana perezi) tissues, meaning detection of this enzyme type also in nonmammalian vertebrates. However, the protein is unique among vertebrate ADHs thus far characterized in having preference for NADP+ rather than NAD+. Similarly, it deviates structurally from other class IV ADHs and has a phylogenetic tree position outside that of the conventional class IV cluster. The NADP+ preference is structurally correlated with a replacement of Asp-223 of all other vertebrate ADHs with Gly-223, largely directing the coenzyme specificity. This residue replacement is expected metabolically to correlate with a change of the reaction direction catalyzed, from preferential alcohol oxidation to preferential aldehyde reduction. This is of importance in cellular growth regulation through retinoic acid formed from retinol/retinal precursors because the enzyme is highly efficient in retinal reduction (kcat/Km = 3.4·104 mm−1min−1). Remaining enzymatic details are also particular but resemble those of the human class I/class IV enzymes. However, overall structural relationships are distant (58–60% residue identity), and residues at substrate binding and coenzyme binding positions are fairly deviant, reflecting the formation of the new activity. The results are concluded to represent early events in the duplicatory origin of the class IV line or of a separate, class IV-type line. In both cases, the novel enzyme illustrates enzymogenesis of classes in the ADH system. The early origin (with tetrapods), the activity (with retinoids), and the specific location of this enzyme (gastric, like the gastric and epithelial location of the human class IV enzyme) suggest important functions of the class IV ADH type in vertebrates.