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PMID: 9546596 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Purification and characterization of the rat liver gamma-butyrobetaine hydroxylase.

Molecular and cellular biochemistry ·Vol. 178 ·No. 1-2 ·1998-01-00 ·Pages 163-8

Galland S, Le Borgne F, Guyonnet D, Clouet P, Demarquoy J

Abstract

The biosynthesis of carnitine from lysine and methionine involves five enzymatic reactions. Gamma-butyrobetaine hydroxylase (BBH; EC 1.14.11.1) is the last enzyme of this pathway. It catalyzes the reaction of hydroxylation of gamma-butyrobetaine to carnitine. This enzyme had never been purified to homogeneity from rat tissue. This paper describes the purification and characterization of the rat liver BBH. This protein has been purified some 413 fold by ion exchange, affinity and gel-filtration chromatographies and appears as a dimere of 43,000 Daltons subunits by PAGE. The affinity chromatography column used in the purification process utilizes 3-(2,2,2-trimethylhydrazinium)propionate (THP), a BBH inhibitor, as the ligand. Polyclonal antibodies were raised against the liver enzyme. They were able to precipitate BBH activity in either a crude liver extract or a purified fraction of the enzyme. Furthermore, it crossreacts with a 43 kDa protein in the liver. No evidence for extra hepatic enzyme was found.

MeSH Terms
Animals Ascorbic Acid/pharmacology Betaine/analogs & derivatives,metabolism Carnitine Catalase/metabolism Catalysis Chromatography, Affinity Enzyme Inhibitors/metabolism Ferrous Compounds/pharmacology Hydroxylation Ketoglutaric Acids/metabolism Kinetics Ligands Liver/enzymology Male Methylhydrazines/metabolism Mixed Function Oxygenases/isolation & purification Molecular Weight Rats Rats, Wistar gamma-Butyrobetaine Dioxygenase
Chemicals
Enzyme Inhibitors Ferrous Compounds Ketoglutaric Acids Ligands Methylhydrazines Betaine gamma-butyrobetaine 3-(2,2,2-trimethylhydrazine)propionate Mixed Function Oxygenases Catalase gamma-Butyrobetaine Dioxygenase Ascorbic Acid Carnitine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Galland S
Université de Bourgogne, Unité de Recherche en Nutrition Cellulaire et Métabolique, Dijon, France.
Le Borgne F
Guyonnet D
Clouet P
Demarquoy J
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Article Info
Journal
Molecular and cellular biochemistry
Abbr.
Mol Cell Biochem
ISSN
0300-8177
DOI
10.1023/a:1006849713407
Published
1998-01-00
Pages
163-8
Language
English
Region
Netherlands
NLM ID
0364456
Subset
IM
External Links
PubMed source DOI: 10.1023/a:1006849713407 View journal details
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