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Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorus lyase pathway.

ABSTRACT: 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.

SUBMITTER: Jochimsen B

PROVIDER: S-EPMC3136323 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature

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Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorus lyase pathway.

Jochimsen Bjarne B Lolle Signe S McSorley Fern R FR Nabi Mariah M Stougaard Jens J Zechel David L DL Hove-Jensen Bjarne B

Proceedings of the National Academy of Sciences of the United States of America 20110624 28

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 polype ...[more]

PMID: 21705661

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Project description:Nutrient elements are important for plant growth. Element stoichiometry considers the balance between different nutrients and how this balance is affected by the environment. So far, focus of plant stoichiometry has mainly been on the three elements carbon (C), nitrogen (N), and phosphorus (P), but many additional elements are essential for proper plant growth. Our overall aim is to test the scaling relations of various additional elements (K, Ca, Mg, S, Cu, Zn, Fe, Mn), by using ten data sets from a range of plant functional types and environmental conditions. To simultaneously handle more than one element, we define a stoichiometric niche volume as the volume of an abstract multidimensional shape in n dimensions, with the n sides of this shape defined by the plant properties in question, here their element concentrations. Thus, a stoichiometric niche volume is here defined as the product of element concentrations. The volumes of N and P (VNP ) are used as the basis, and we investigate how the volume of other elements (VOth ) scales with respect to VNP¸ with the intention to explore if the concentrations of other elements increase faster (scaling exponent > 1) or slower (<1) than the concentrations of N and P. For example, scaling exponents >1 suggest that favorable conditions for plant growth, i.e., environments rich in N and P, may require proportionally higher uptake of other essential elements than poor conditions. We show that the scaling exponent is rather insensitive to environmental conditions or plant species, and ranges from 0.900 to 2.479 (average 1.58) in nine out of ten data sets. For single elements, Mg has the smallest scaling exponent (0.031) and Mn the largest (2.147). Comparison between laboratory determined stoichiometric relations and field observations suggest that element uptake in field conditions often exceeds the minimal physiological requirements. The results provide evidence for the view that the scaling relations previously reported for N and P can be extended to other elements; and that N and P are the driving elements in plant stoichiometric relations. The stoichiometric niche volumes defined here could be used to predict plant performances in different environments.

Dataset Information

Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorus lyase pathway.

Publications

Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorus lyase pathway.

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