Crotonobetaine reductase from Escherichia coli consists of two proteins.
Crotonobetaine reductase from Escherichia coli is composed of two proteins (component I (CI) and component II (CII)). CI has been purified to electrophoretic homogeneity from a cell-free extract of E. coli O44 K74. The purified protein shows l(-)-carnitine dehydratase activity and its N-terminal amino acid sequence is identical to the caiB gene product from E. coli O44 K74. The relative molecular mass of CI has been determined to be 86100. It is composed of two identical subunits with a molecular mass of 42600. The isoelectric point of CI was found to be 4.3. CII was purified from an overexpression strain in one step by ion exchange chromatography on Fractogel EMD TMAE 650(S). The N-terminal amino acid sequence of CII shows absolute identity with the N-terminal sequence of the caiA gene product, i.e. of the postulated crotonobetaine reductase. The relative molecular mass of the protein is 164400 and it is composed of four identical subunits of molecular mass 41500. The isoelectric point of CII is 5.6. CII contains non-covalently bound FAD in a molar ratio of 1:1. In the crotonobetaine reductase reaction one dimer of CI associates with one tetramer of CII. A still unknown low-molecular-mass effector described for the l(-)-carnitine dehydratase is also necessary for crotonobetaine reductase activity. Monoclonal antibodies were raised against the two components of crotonobetaine reductase.
Preusser
A
A
Institut für Biochemie, Fakultät für Biowissenschaften, Pharmazie und Psychologie, Universität Leipzig, Talstr. 33, D-04103, Leipzig, Germany.
Wagner
U
U
Elssner
T
T
Kleber
H P
HP
D000217:Acyltransferases; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001622:Betaine / Q000031:analogs & derivatives / Q000737:chemistry; D002331:Carnitine; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D006836:Hydro-Lyases / Q000737:chemistry; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000276:immunology; D010088:Oxidoreductases / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D001622:Betaine; C002889:gamma-butyrobetaine; C091403:CaiA protein, E coli; D010088:Oxidoreductases; C067153:crotonobetaine reductase; D000217:Acyltransferases; C060278:crotonobetainyl-CoA carnitine coenzyme A transferase; D006836:Hydro-Lyases; D002331:Carnitine