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Substrate specificities of 3-oxoacyl-CoA thiolase A and sterol carrier protein 2/3-oxoacyl-CoA thiolase purified from normal rat liver peroxisomes. Sterol carrier protein 2/3-oxoacyl-CoA thiolase is involved in the metabolism of 2-methyl-branched fatty acids and bile acid intermediates.
J Biol Chem. 1997 Oct 10; 272(41):26023-31.JB

Abstract

The two main thiolase activities present in isolated peroxisomes from normal rat liver were purified to near homogeneity. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the first enzyme preparation displayed a single band of 41 kDa that was identified as 3-oxoacyl-CoA thiolase A (thiolase A) by N-terminal amino acid sequencing. The second enzyme preparation consisted of a 58- and a 46-kDa band. The 58-kDa polypeptide reacted with antibodies raised against either sterol carrier protein 2 or the thiolase domain of sterol carrier protein 2/3-oxoacyl-CoA thiolase (SCP-2/thiolase), formerly also called sterol carrier protein X, whereas the 46-kDa polypeptide reacted only with the antibodies raised against the thiolase domain. Internal peptide sequencing confirmed that the 58-kDa polypeptide is SCP-2/thiolase and that the 46-kDa polypeptide is the thiolase domain of SCP-2/thiolase. Thiolase A catalyzed the cleavage of short, medium, and long straight chain 3-oxoacyl-CoAs, medium chain 3-oxoacyl-CoAs being the best substrates. The enzyme was inactive with the 2-methyl-branched 3-oxo-2-methylpalmitoyl-CoA and with the bile acid intermediate 24-oxo-trihydroxycoprostanoyl-CoA. SCP-2/thiolase was active with medium and long straight chain 3-oxoacyl-CoAs but also with the 2-methyl-branched 3-oxoacyl-CoA and the bile acid intermediate. In peroxisomal extracts, more than 90% of the thiolase activity toward straight chain 3-oxoacyl-CoAs was associated with thiolase A. Kinetic parameters (Km and Vmax) were determined for each enzyme with the different substrates. Our results indicate the following: 1) the two (main) thiolases present in peroxisomes from normal rat liver are thiolase A and SCP-2/thiolase; 2) thiolase A is responsible for the thiolytic cleavage of straight chain 3-oxoacyl-CoAs; and 3) SCP-2/thiolase is responsible for the thiolytic cleavage of the 3-oxoacyl-CoA derivatives of 2-methyl-branched fatty acids and the side chain of cholesterol.

Authors+Show Affiliations

Katholieke Universiteit Leuven, Departement Moleculaire Celbiologie, Afdeling, Campus Gasthuisberg (O & N), Herestraat 49, B-3000 Leuven, Belgium.No 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

9325339

Citation

Antonenkov, V D., et al. "Substrate Specificities of 3-oxoacyl-CoA Thiolase a and Sterol Carrier Protein 2/3-oxoacyl-CoA Thiolase Purified From Normal Rat Liver Peroxisomes. Sterol Carrier Protein 2/3-oxoacyl-CoA Thiolase Is Involved in the Metabolism of 2-methyl-branched Fatty Acids and Bile Acid Intermediates." The Journal of Biological Chemistry, vol. 272, no. 41, 1997, pp. 26023-31.
Antonenkov VD, Van Veldhoven PP, Waelkens E, et al. Substrate specificities of 3-oxoacyl-CoA thiolase A and sterol carrier protein 2/3-oxoacyl-CoA thiolase purified from normal rat liver peroxisomes. Sterol carrier protein 2/3-oxoacyl-CoA thiolase is involved in the metabolism of 2-methyl-branched fatty acids and bile acid intermediates. J Biol Chem. 1997;272(41):26023-31.
Antonenkov, V. D., Van Veldhoven, P. P., Waelkens, E., & Mannaerts, G. P. (1997). Substrate specificities of 3-oxoacyl-CoA thiolase A and sterol carrier protein 2/3-oxoacyl-CoA thiolase purified from normal rat liver peroxisomes. Sterol carrier protein 2/3-oxoacyl-CoA thiolase is involved in the metabolism of 2-methyl-branched fatty acids and bile acid intermediates. The Journal of Biological Chemistry, 272(41), 26023-31.
Antonenkov VD, et al. Substrate Specificities of 3-oxoacyl-CoA Thiolase a and Sterol Carrier Protein 2/3-oxoacyl-CoA Thiolase Purified From Normal Rat Liver Peroxisomes. Sterol Carrier Protein 2/3-oxoacyl-CoA Thiolase Is Involved in the Metabolism of 2-methyl-branched Fatty Acids and Bile Acid Intermediates. J Biol Chem. 1997 Oct 10;272(41):26023-31. PubMed PMID: 9325339.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Substrate specificities of 3-oxoacyl-CoA thiolase A and sterol carrier protein 2/3-oxoacyl-CoA thiolase purified from normal rat liver peroxisomes. Sterol carrier protein 2/3-oxoacyl-CoA thiolase is involved in the metabolism of 2-methyl-branched fatty acids and bile acid intermediates. AU - Antonenkov,V D, AU - Van Veldhoven,P P, AU - Waelkens,E, AU - Mannaerts,G P, PY - 1997/11/5/pubmed PY - 1997/11/5/medline PY - 1997/11/5/entrez SP - 26023 EP - 31 JF - The Journal of biological chemistry JO - J Biol Chem VL - 272 IS - 41 N2 - The two main thiolase activities present in isolated peroxisomes from normal rat liver were purified to near homogeneity. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the first enzyme preparation displayed a single band of 41 kDa that was identified as 3-oxoacyl-CoA thiolase A (thiolase A) by N-terminal amino acid sequencing. The second enzyme preparation consisted of a 58- and a 46-kDa band. The 58-kDa polypeptide reacted with antibodies raised against either sterol carrier protein 2 or the thiolase domain of sterol carrier protein 2/3-oxoacyl-CoA thiolase (SCP-2/thiolase), formerly also called sterol carrier protein X, whereas the 46-kDa polypeptide reacted only with the antibodies raised against the thiolase domain. Internal peptide sequencing confirmed that the 58-kDa polypeptide is SCP-2/thiolase and that the 46-kDa polypeptide is the thiolase domain of SCP-2/thiolase. Thiolase A catalyzed the cleavage of short, medium, and long straight chain 3-oxoacyl-CoAs, medium chain 3-oxoacyl-CoAs being the best substrates. The enzyme was inactive with the 2-methyl-branched 3-oxo-2-methylpalmitoyl-CoA and with the bile acid intermediate 24-oxo-trihydroxycoprostanoyl-CoA. SCP-2/thiolase was active with medium and long straight chain 3-oxoacyl-CoAs but also with the 2-methyl-branched 3-oxoacyl-CoA and the bile acid intermediate. In peroxisomal extracts, more than 90% of the thiolase activity toward straight chain 3-oxoacyl-CoAs was associated with thiolase A. Kinetic parameters (Km and Vmax) were determined for each enzyme with the different substrates. Our results indicate the following: 1) the two (main) thiolases present in peroxisomes from normal rat liver are thiolase A and SCP-2/thiolase; 2) thiolase A is responsible for the thiolytic cleavage of straight chain 3-oxoacyl-CoAs; and 3) SCP-2/thiolase is responsible for the thiolytic cleavage of the 3-oxoacyl-CoA derivatives of 2-methyl-branched fatty acids and the side chain of cholesterol. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/9325339/Substrate_specificities_of_3_oxoacyl_CoA_thiolase_A_and_sterol_carrier_protein_2/3_oxoacyl_CoA_thiolase_purified_from_normal_rat_liver_peroxisomes__Sterol_carrier_protein_2/3_oxoacyl_CoA_thiolase_is_involved_in_the_metabolism_of_2_methyl_branched_fatty_acids_and_bile_acid_intermediates_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(18)60208-X DB - PRIME DP - Unbound Medicine ER -