Reconstitution of glucose uptake and chemotaxis in Pseudomonas aeruginosa glucose transport defective mutants

1993 ◽  
Vol 39 (11) ◽  
pp. 1079-1083 ◽  
Author(s):  
Laura M. Sly ◽  
Elizabeth A. Worobec ◽  
Richard E. Perkins ◽  
Paul V. Phibbs Jr.
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Glucose Uptake ◽  
Glucose Transport ◽  
Binding Protein ◽  
Cell Uptake ◽  
Wild Type ◽  
Binding Assays ◽  
Glucose Binding ◽  
Glucose Transport System ◽  
Glucose Binding Protein

Wild-type glucose uptake and glucose chemotaxis activities were restored in glucose transport defective Pseudomonas aeruginosa strains PFB360 and PFB362 after introduction of plasmid pPZ129, containing a 1.1-kilobase DNA fragment that is essential for the expression of the P. aeruginosa periplasmic glucose binding protein. The restoration of glucose uptake and chemotaxis to wild-type levels in these strains was also achieved by reconstitution with cold-shock fluid and purified glucose binding protein isolated from P. aeruginosa PA01 wild-type strain H103 grown in conditions resulting in the induction of the high-affinity glucose transport system. Glucose uptake was determined by whole cell uptake and shock fluid binding of D-[U-14C]glucose, using standard filter binding assays. Positive chemotaxis towards glucose was assessed by capillary assays using 10 mM glucose, the amount required for optimal chemotaxis, and judged by plating capillary contents accumulated after 30 min.Key words: glucose-binding protein, Pseudomonas aeruginosa, glucose transport, chemotaxis.

Substrate specificity of the high-affinity glucose transport system of Pseudomonas aeruginosa

1993 ◽  
Vol 39 (7) ◽  
pp. 722-725 ◽  
Author(s):  
John L. Wylie ◽  
Elizabeth A. Worobec
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Glucose Transport ◽  
Transport System ◽  
Binding Protein ◽  
High Affinity ◽  
Glucose Binding ◽  
Glucose Transport System ◽  
Glucose Binding Protein

Specificity of the high-affinity glucose transport system of Pseudomonas aeruginosa was examined. At a concentration of [14C]glucose near the Vmax of the system, inhibition by maltose, galactose, and xylose was detected. This inhibition is similar to that detected in earlier in vivo studies and correlates with the known specificity of OprB, a glucose-specific porin of P. aeruginosa. At a level of [14C]glucose 100 times lower, only unlabelled glucose inhibited uptake to any extent. This matches the known in vitro specificity of the periplasmic glucose binding protein. These findings were used to explain the discrepancy between earlier in vivo and in vitro results reported in the literature.Key words: Pseudomonas aeruginosa, glucose transport, OprB, glucose binding protein.

Labeling of a glucose binding protein in the rabbit intestinal brush border membrane

1978 ◽  
Vol 56 (5) ◽  
pp. 760-770 ◽  
Author(s):  
J. Lemaire ◽  
D. Maestracci
Keyword(s):  
Molecular Weight ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Binding Protein ◽  
Electrophoretic Pattern ◽  
Protein Band ◽  
Relative Mobility ◽  
Intestinal Brush Border ◽  
Glucose Binding ◽  
Glucose Binding Protein

Using a double labeling method based on the method of Thomas (Thomas, L. 1973. Isolation of N-ethylmaleimide-labeled phloridzin-sensitive D-glucose binding protein of brash border membrane from rat kidney cortex. Biochim. Biophys. Acta, 291, 454–464.), with radioactive N-ethylmaleimide ([3H]NEM and [14C]NEM) in the presence and absence of D-glucose, a protein band which is periodic acid – Schiff staining insensitive and which has a relative mobility (Rm) of 0.55 (corresponding to a molecular weight of 51 000 daltons) as determined by sodium dodecyl sulfate (SDS) electrophoresis was labeled preferentially.When radioactive p-hydroxymercuriphenylsulfonate ([203Hg]PCMBS) is used in the presence and absence of D-glucose, as described by Smith et al. (SMITH, M. W., FERGUSON, D. R., and BURTON, K. A. 1975. Glucose- and phloridzin-protected thiol groups in pig intestinal brush border membranes. Biochern. J. 147, 617–619.), a protein band which has a relative mobility of 0.62 and a corresponding molecular weight of 42 000 daltons was labeled.Control experiments have shown that increasing concentrations of nonradioactive NEM (0.1–5.0 mM) do not substantially modify the electrophoretic pattern of SDS-solubilized brush border membrane. Nonradioactive PCMBS (0.1–10 mM), on the other hand, modifies the electrophoretic pattern and especially causes a change in relative mobility of the 0.55 protein band which migrates after 1 mM PCMBS treatment with a Rm of 0.62.The effect of 1 mM PCMBS can be reversed by adding L-cysteine or dithiotreitol.Actin extracted from rabbit muscle migrates with the same Rm as the 0.55 protein band in our electrophoretic conditions.

Pressure effect on the stability and the conformational dynamics of the D-Galactose/D-Glucose-binding protein from Escherichia coli

2005 ◽  
Vol 62 (1) ◽  
pp. 193-201 ◽  
Author(s):  
Anna Marabotti ◽  
Petr Herman ◽  
Maria Staiano ◽  
Antonio Varriale ◽  
Marcella de Champdoré ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pressure Effect ◽  
Binding Protein ◽  
Conformational Dynamics ◽  
Glucose Binding ◽  
The Stability ◽  
Glucose Binding Protein

Preformulation Characterization, Stabilization, and Formulation Design for the Acrylodan-Labeled Glucose-Binding Protein SM4-AC

2017 ◽  
Vol 106 (5) ◽  
pp. 1197-1210 ◽  
Author(s):  
Neha Sahni ◽  
Rajoshi Chaudhuri ◽  
John M. Hickey ◽  
Prakash Manikwar ◽  
Ajit D’Souza ◽  
...  
Keyword(s):  
Binding Protein ◽  
Formulation Design ◽  
Glucose Binding ◽  
Glucose Binding Protein

scholarly journals Structure of a periplasmic glucose-binding protein fromThermotoga maritima

2012 ◽  
Vol 68 (12) ◽  
pp. 1460-1464 ◽  
Author(s):  
Kandavelu Palani ◽  
Desigan Kumaran ◽  
Stephen K. Burley ◽  
Subramanyam Swaminathan
Keyword(s):  
Binding Protein ◽  
Glucose Binding ◽  
Glucose Binding Protein
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