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Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis.
Biochem J. 1997 Dec 01; 328 (Pt 2):377-82.BJ

Abstract

Rat liver peroxisomes contain two multifunctional enzymes: (1) perMFE-1 [2-enoyl-CoA hydratase 1/Delta3,Delta2-enoyl-CoA isomerase/(S)-3-hydroxyacyl-CoA dehydrogenase] and (2) perMFE-2 [2-enoyl-CoA hydratase 2/(R)-3-hydroxyacyl-CoA dehydrogenase]. To investigate the role of the hydratase activity of perMFE-2 in beta-oxidation, a truncated version of perMFE-2 was expressed in Escherichia coli as a recombinant protein. The protein catalyses the hydration of straight-chain (2E)-enoyl-CoAs to (3R)-hydroxyacyl-CoAs, but it is devoid of hydratase 1 [(2E)-enoyl-CoA to (3S)-hydroxyacyl-CoA] and (3R)-hydroxyacyl-CoA dehydrogenase activities. The purified enzyme (46 kDa hydratase 2) can be stored as an active enzyme for at least half a year. The recombinant enzyme hydrates (24E)-3alpha,7alpha,12alpha-trihydroxy- 5beta-cholest-24-enoyl-CoA to (24R,25R)-3alpha,7alpha,12alpha, 24-tetrahydroxy-5beta-cholestanoyl-CoA, which has previously been characterized as a physiological intermediate in bile acid synthesis. The stereochemistry of the products indicates that the hydration reaction catalysed by the enzyme proceeds via a syn mechanism. A monofunctional 2-enoyl-CoA hydratase 2 has not been observed as a wild-type protein. The recombinant 46 kDa hydratase 2 described here survives in a purified form under storage, thus being the first protein of this type amenable to application as a tool in metabolic studies.

Authors+Show Affiliations

Biocenter Oulu, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9371691

Citation

Qin, Y M., et al. "Recombinant 2-enoyl-CoA Hydratase Derived From Rat Peroxisomal Multifunctional Enzyme 2: Role of the Hydratase Reaction in Bile Acid Synthesis." The Biochemical Journal, vol. 328 (Pt 2), 1997, pp. 377-82.
Qin YM, Haapalainen AM, Conry D, et al. Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis. Biochem J. 1997;328 (Pt 2):377-82.
Qin, Y. M., Haapalainen, A. M., Conry, D., Cuebas, D. A., Hiltunen, J. K., & Novikov, D. K. (1997). Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis. The Biochemical Journal, 328 (Pt 2), 377-82.
Qin YM, et al. Recombinant 2-enoyl-CoA Hydratase Derived From Rat Peroxisomal Multifunctional Enzyme 2: Role of the Hydratase Reaction in Bile Acid Synthesis. Biochem J. 1997 Dec 1;328 (Pt 2):377-82. PubMed PMID: 9371691.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis. AU - Qin,Y M, AU - Haapalainen,A M, AU - Conry,D, AU - Cuebas,D A, AU - Hiltunen,J K, AU - Novikov,D K, PY - 1998/1/24/pubmed PY - 1998/1/24/medline PY - 1998/1/24/entrez SP - 377 EP - 82 JF - The Biochemical journal JO - Biochem J VL - 328 (Pt 2) N2 - Rat liver peroxisomes contain two multifunctional enzymes: (1) perMFE-1 [2-enoyl-CoA hydratase 1/Delta3,Delta2-enoyl-CoA isomerase/(S)-3-hydroxyacyl-CoA dehydrogenase] and (2) perMFE-2 [2-enoyl-CoA hydratase 2/(R)-3-hydroxyacyl-CoA dehydrogenase]. To investigate the role of the hydratase activity of perMFE-2 in beta-oxidation, a truncated version of perMFE-2 was expressed in Escherichia coli as a recombinant protein. The protein catalyses the hydration of straight-chain (2E)-enoyl-CoAs to (3R)-hydroxyacyl-CoAs, but it is devoid of hydratase 1 [(2E)-enoyl-CoA to (3S)-hydroxyacyl-CoA] and (3R)-hydroxyacyl-CoA dehydrogenase activities. The purified enzyme (46 kDa hydratase 2) can be stored as an active enzyme for at least half a year. The recombinant enzyme hydrates (24E)-3alpha,7alpha,12alpha-trihydroxy- 5beta-cholest-24-enoyl-CoA to (24R,25R)-3alpha,7alpha,12alpha, 24-tetrahydroxy-5beta-cholestanoyl-CoA, which has previously been characterized as a physiological intermediate in bile acid synthesis. The stereochemistry of the products indicates that the hydration reaction catalysed by the enzyme proceeds via a syn mechanism. A monofunctional 2-enoyl-CoA hydratase 2 has not been observed as a wild-type protein. The recombinant 46 kDa hydratase 2 described here survives in a purified form under storage, thus being the first protein of this type amenable to application as a tool in metabolic studies. SN - 0264-6021 UR - https://www.unboundmedicine.com/medline/citation/9371691/Recombinant_2_enoyl_CoA_hydratase_derived_from_rat_peroxisomal_multifunctional_enzyme_2:_role_of_the_hydratase_reaction_in_bile_acid_synthesis_ L2 - https://portlandpress.com/biochemj/article-lookup/doi/10.1042/bj3280377 DB - PRIME DP - Unbound Medicine ER -