Journal article
The hemA gene encoding glutamyl-tRNA reductase from the archaeon Methanobacterium thermoautotrophicum strain Marburg
Bioorganic & medicinal chemistry, Vol.4(7), pp.1089-1095
1996
DOI: 10.1016/0968-0896(96)00098-3
PMID: 8831980
Abstract
In archaea the first general tetrapyrrole precursor 5-aminolevulinic acid (ALA) is formed via the tRNA-dependent five-carbon pathway from glutamate. We have cloned the
hemA gene encoding the central enzyme of the pathway glutamyl- tRNA reductase from the methanogenic archacon
Methanobacterium thermoautotrophicum by complementation of an
Escherichia coli hemA mutant to ALA prototrophy. An 1194 bp open reading frame that encodes a 398 amino acid polypeptide with the calculated
M
r
44,509 was detected. The deduced amino acid sequence showed 20–35% amino acid identity to bacterial HemAs with the highest identity score to the
Pseudomonas aeruginosa HemA. An identity of approximately 22% was found to plant HemAs. Glutamyl-tRNA reductase activity was shown for the
M. thermoautotrophicum HemA after overexpression in
E. coli and partial purification. The enzymatic reaction catalyzed by the partially purified enzyme revealed a temperature optimum of 65 °C at an optimal pH of 7.0. The reductase utilized preferentially NADPH for the reduction of the activated carboxyl group. The presence of ATP and GTP showed no obvious influence on catalysis.
The cloning and characterization of the
hemA gene from
Methanobacterium thermoautotrophicum encoding glutamyl tRNA reductase is reported. An initial biochemical characterization of the recombinant HemA protein is described.
Details
- Title: Subtitle
- The hemA gene encoding glutamyl-tRNA reductase from the archaeon Methanobacterium thermoautotrophicum strain Marburg
- Creators
- Christoph Hungerer - Laboratorium für Mikrobiologie des Fachbereich Biologie der Philipps-Universität and Max-Planck-Institut für Terrestrische Mikrobiologie, Karl-von-Frisch-Strasse, 35043 Marburg, GermanyDavid S Weiss - Harvard Medical School, Department of Microbiology, 200 Longwood Avenue, Boston, MA 02115, U.S.ARudolf K Thauer - Laboratorium für Mikrobiologie des Fachbereich Biologie der Philipps-Universität and Max-Planck-Institut für Terrestrische Mikrobiologie, Karl-von-Frisch-Strasse, 35043 Marburg, GermanyDieter Jahn - Laboratorium für Mikrobiologie des Fachbereich Biologie der Philipps-Universität and Max-Planck-Institut für Terrestrische Mikrobiologie, Karl-von-Frisch-Strasse, 35043 Marburg, Germany
- Resource Type
- Journal article
- Publication Details
- Bioorganic & medicinal chemistry, Vol.4(7), pp.1089-1095
- Publisher
- Elsevier Ltd
- DOI
- 10.1016/0968-0896(96)00098-3
- PMID
- 8831980
- ISSN
- 0968-0896
- eISSN
- 1464-3391
- Language
- English
- Date published
- 1996
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984001114502771
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