Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorus lyase pathway
| Autor: | Bjarne Jochimsen, Bjarne Hove-Jensen, Jens Stougaard, David L. Zechel, Mariah Nabi, Signe Lolle, Fern R. McSorley |
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| Rok vydání: | 2011 |
| Předmět: |
DNA
Bacterial Operon Protein subunit Organophosphonates Lyases Biology medicine.disease_cause Models Biological Regulon 03 medical and health sciences chemistry.chemical_compound Affinity chromatography Multienzyme Complexes Escherichia coli medicine Cloning Molecular Sequence Deletion 030304 developmental biology 0303 health sciences Multidisciplinary Base Sequence Molecular mass 030306 microbiology Biological Sciences Lyase Molecular Weight Protein Subunits Genes Biochemistry chemistry Genes Bacterial Metabolic Networks and Pathways DNA |
| Zdroj: | Proceedings of the National Academy of Sciences Jochimsen, B, Lolle, S, McSorley, F, Nabi, M, Stougaard, J, Zechel, D & Hove-Jensen, B 2011, ' Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorous lyase pathway ', Proceedings of the National Academy of Sciences of the United States of America, vol. 108, no. 28, pp. 11393-8 . |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1104922108 |
| Popis: | Organophosphonate utilization by Escherichia coli requires the 14 cistrons of the phnCDEFGHIJKLMNOP operon, of which the carbon-phosphorus lyase has been postulated to consist of the seven polypeptides specified by phnG to phnM . A 5,660-bp DNA fragment encompassing phnGHIJKLM is cloned, followed by expression in E. coli and purification of Phn-polypeptides. PhnG, PhnH, PhnI, PhnJ, and PhnK copurify as a protein complex by ion-exchange, size-exclusion, and affinity chromatography. The five polypeptides also comigrate in native-PAGE. Cross-linking of the purified protein complex reveals a close proximity of PhnG, PhnI, PhnJ, and PhnK, as these subunits disappear concomitant with the formation of large cross-linked protein complexes. Two molecular forms are identified, a major form of molecular mass of approximately 260 kDa, a minor form of approximately 640 kDa. The stoichiometry of the protein complex is suggested to be PhnG 4 H 2 I 2 J 2 K. Deletion of individual phn genes reveals that a strain harboring plasmid-borne phnGHIJ produces a protein complex consisting of PhnG, PhnH, PhnI, and PhnJ, whereas a strain harboring plasmid-borne phnGIJK produces a protein complex consisting of PhnG and PhnI. We conclude that phnGHIJK specify a soluble multisubunit protein complex essential for organophosphonate utilization. |
| Databáze: | OpenAIRE |
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