Purification and properties of glucose 1-phosphate binding protein in Agrobacterium tumefaciens

The secreted protein pattern of Streptomyces lividans depends on the carbon source present in the culture media. One protein that shows the most dramatic change is the high-affinity phosphate-binding protein PstS, which is strongly accumulated in the supernatant of liquid cultures containing high concentrations (>3 %) of certain sugars, such as fructose, galactose and mannose. The promoter region of this gene and that of its Streptomyces coelicolor homologue were used to drive the expression of a xylanase in S. lividans that was accumulated in the culture supernatant when grown in the presence of fructose. PstS accumulation was dramatically increased in a S. lividans polyphosphate kinase null mutant (Δppk) and was impaired in a deletion mutant lacking phoP, the transcriptional regulator gene of the two-component phoR-phoP system that controls the Pho regulon. Deletion of the pstS genes in S. lividans and S. coelicolor impaired phosphate transport and accelerated differentiation and sporulation on solid media. Complementation with a single copy in a S. lividans pstS null mutant returned phosphate transport and sporulation to levels similar to those of the wild-type strain. The present work demonstrates that carbon and phosphate metabolism are linked in the regulation of genes and that this can trigger the genetic switch towards morphogenesis.

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A Dynamical Investigation of Acrylodan-Labeled Mutant Phosphate Binding Protein

Analytical Chemistry ◽

10.1021/ac980800g ◽

1999 ◽

Vol 71 (3) ◽

pp. 589-595 ◽

Cited By ~ 13

Author(s):

Jeffrey S. Lundgren ◽

Lyndon L. E. Salins ◽

Irina Kaneva ◽

Sylvia Daunert

Keyword(s):

Binding Protein ◽

Phosphate Binding ◽

Phosphate Binding Protein

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Preliminary time-of-flight neutron diffraction studies ofEscherichia coliABC transport receptor phosphate-binding protein at the Protein Crystallography Station

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x14009704 ◽

2014 ◽

Vol 70 (6) ◽

pp. 819-822 ◽

Cited By ~ 3

Author(s):

K. H. Sippel ◽

J. Bacik ◽

F. A. Quiocho ◽

S. Z. Fisher

Keyword(s):

Neutron Diffraction ◽

Binding Protein ◽

Protein Crystallography ◽

Phosphate Transport ◽

Phosphate Binding ◽

Science Center ◽

Data Set ◽

Acceptor Group ◽

Phosphate Binding Protein ◽

Ph Range

Inorganic phosphate is an essential molecule for all known life. Organisms have developed many mechanisms to ensure an adequate supply, even in low-phosphate conditions. In prokaryotes phosphate transport is instigated by the phosphate-binding protein (PBP), the initial receptor for the ATP-binding cassette (ABC) phosphate transporter. In the crystal structure of the PBP–phosphate complex, the phosphate is completely desolvated and sequestered in a deep cleft and is bound by 13 hydrogen bonds: 12 to protein NH and OH donor groups and one to a carboxylate acceptor group. The carboxylate plays a key recognition role by accepting a phosphate hydrogen. PBP phosphate affinity is relatively consistent across a broad pH range, indicating the capacity to bind monobasic (H2PO4−) and dibasic (HPO42−) phosphate; however, the mechanism by which it might accommodate the second hydrogen of monobasic phosphate is unclear. To answer this question, neutron diffraction studies were initiated. Large single crystals with a volume of 8 mm3were grown and subjected to hydrogen/deuterium exchange. A 2.5 Å resolution data set was collected on the Protein Crystallography Station at the Los Alamos Neutron Science Center. Initial refinement of the neutron data shows significant nuclear density, and refinement is ongoing. This is the first report of a neutron study from this superfamily.

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