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Role of aspartic acid 38 in the cofactor specificity of Drosophila alcohol dehydrogenase.
Eur J Biochem. 1991 Dec 05; 202(2):263-7.EJ

Abstract

Drosophila alcohol dehydrogenase (ADH), an NAD(+)-dependent dehydrogenase, shares little sequence similarity with horse liver ADH. However, these two enzymes do have substantial similarity in their secondary structure at the NAD(+)-binding domain [Benyajati, C., Place, A. P., Powers, D. A. & Sofer, W. (1981) Proc. Natl Acad. Sci. USA 78, 2717-2721]. Asp38, a conserved residue between Drosophila and horse liver ADH, appears to interact with the hydroxyl groups of the ribose moiety in the AMP portion of NAD+. A secondary-structure comparison between the nucleotide-binding domain of NAD(+)-dependent enzymes and that of NADP(+)-dependent enzymes also suggests that Asp38 could play an important role in cofactor specificity. Mutating Asp38 of Drosophila ADH into Asn38 decreases Km(app)NADP 62-fold and increases kcat/Km(app)NADP 590-fold at pH 9.8, when compared with wild-type ADH. These results suggest that Asp38 is in the NAD(+)-binding domain and its substituent, Asn38, allows Drosophila ADH to use both NAD+ and NADP+ as its cofactor. The observations from the experiments of thermal denaturation and kinetic measurement with pH also confirm that the repulsion between the negative charges of Asp38 and 2'-phosphate of NADP+ is the major energy barrier for NADP+ to serve as a cofactor for Drosophila ADH.

Authors+Show Affiliations

Department of Biochemistry, Louisiana State University, Baton Rouge 70803.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

1761031

Citation

Chen, Z, et al. "Role of Aspartic Acid 38 in the Cofactor Specificity of Drosophila Alcohol Dehydrogenase." European Journal of Biochemistry, vol. 202, no. 2, 1991, pp. 263-7.
Chen Z, Lee WR, Chang SH. Role of aspartic acid 38 in the cofactor specificity of Drosophila alcohol dehydrogenase. Eur J Biochem. 1991;202(2):263-7.
Chen, Z., Lee, W. R., & Chang, S. H. (1991). Role of aspartic acid 38 in the cofactor specificity of Drosophila alcohol dehydrogenase. European Journal of Biochemistry, 202(2), 263-7.
Chen Z, Lee WR, Chang SH. Role of Aspartic Acid 38 in the Cofactor Specificity of Drosophila Alcohol Dehydrogenase. Eur J Biochem. 1991 Dec 5;202(2):263-7. PubMed PMID: 1761031.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Role of aspartic acid 38 in the cofactor specificity of Drosophila alcohol dehydrogenase. AU - Chen,Z, AU - Lee,W R, AU - Chang,S H, PY - 1991/12/5/pubmed PY - 1991/12/5/medline PY - 1991/12/5/entrez SP - 263 EP - 7 JF - European journal of biochemistry JO - Eur. J. Biochem. VL - 202 IS - 2 N2 - Drosophila alcohol dehydrogenase (ADH), an NAD(+)-dependent dehydrogenase, shares little sequence similarity with horse liver ADH. However, these two enzymes do have substantial similarity in their secondary structure at the NAD(+)-binding domain [Benyajati, C., Place, A. P., Powers, D. A. & Sofer, W. (1981) Proc. Natl Acad. Sci. USA 78, 2717-2721]. Asp38, a conserved residue between Drosophila and horse liver ADH, appears to interact with the hydroxyl groups of the ribose moiety in the AMP portion of NAD+. A secondary-structure comparison between the nucleotide-binding domain of NAD(+)-dependent enzymes and that of NADP(+)-dependent enzymes also suggests that Asp38 could play an important role in cofactor specificity. Mutating Asp38 of Drosophila ADH into Asn38 decreases Km(app)NADP 62-fold and increases kcat/Km(app)NADP 590-fold at pH 9.8, when compared with wild-type ADH. These results suggest that Asp38 is in the NAD(+)-binding domain and its substituent, Asn38, allows Drosophila ADH to use both NAD+ and NADP+ as its cofactor. The observations from the experiments of thermal denaturation and kinetic measurement with pH also confirm that the repulsion between the negative charges of Asp38 and 2'-phosphate of NADP+ is the major energy barrier for NADP+ to serve as a cofactor for Drosophila ADH. SN - 0014-2956 UR - https://www.unboundmedicine.com/medline/citation/1761031/Role_of_aspartic_acid_38_in_the_cofactor_specificity_of_Drosophila_alcohol_dehydrogenase_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0014-2956&date=1991&volume=202&issue=2&spage=263 DB - PRIME DP - Unbound Medicine ER -