scholarly journals Cellobiose Uptake in the Hyperthermophilic ArchaeonPyrococcus furiosus Is Mediated by an Inducible, High-Affinity ABC Transporter

2001 ◽  
Vol 183 (17) ◽  
pp. 4979-4984 ◽  
Author(s):  
Sonja M. Koning ◽  
Marieke G. L. Elferink ◽  
Wil N. Konings ◽  
Arnold J. M. Driessen
Keyword(s):  
Escherichia Coli ◽  
Gene Cluster ◽  
Abc Transporter ◽  
Transport System ◽  
Abc Transporters ◽  
Pyrococcus Furiosus ◽  
Binding Protein ◽  
Hyperthermophilic Archaeon ◽  
High Affinity ◽  
Dependent Transport

ABSTRACT The hyperthermophilic archaeon Pyrococcus furiosuscan utilize different β-glucosides, like cellobiose and laminarin. Cellobiose uptake occurs with high affinity (K m = 175 nM) and involves an inducible binding protein-dependent transport system. The cellobiose binding protein (CbtA) was purified from P. furiosusmembranes to homogeneity as a 70-kDa glycoprotein. CbtA not only binds cellobiose but also cellotriose, cellotetraose, cellopentaose, laminaribiose, laminaritriose, and sophorose. The cbtAgene was cloned and functionally expressed in Escherichia coli. cbtA belongs to a gene cluster that encodes a transporter that belongs to the Opp family of ABC transporters.

scholarly journals Biochemical evidence for the presence of two α-glucoside ABC-transport systems in the hyperthermophilic archaeonPyrococcus furiosus

Archaea ◽  
2002 ◽  
Vol 1 (1) ◽  
pp. 19-25 ◽  
Author(s):  
Sonja M. Koning ◽  
Wil N. Konings ◽  
Arnold J.M. Driessen
Keyword(s):  
Transport System ◽  
Pyrococcus Furiosus ◽  
Binding Protein ◽  
Transport Systems ◽  
Atp Binding ◽  
Hyperthermophilic Archaeon ◽  
Biochemical Evidence ◽  
Abc Transport ◽  
Maltose Transporter ◽  
Atp Binding Cassette

The hyperthermophilic archaeonPyrococcus furiosuscan utilize different carbohydrates, such as starch, maltose and trehalose. Uptake of α-glucosides is mediated by two different, binding protein-dependent, ATP-binding cassette (ABC)-type transport systems. The maltose transporter also transports trehalose, whereas the maltodextrin transport system mediates the uptake of maltotriose and higher malto-oligosaccharides, but not maltose. Both transport systems are induced during growth on their respective substrates.

scholarly journals Noncanonical role for the binding protein in substrate uptake by the MetNI methionine ATP Binding Cassette (ABC) transporter

2018 ◽  
Vol 115 (45) ◽  
pp. E10596-E10604 ◽  
Author(s):  
Phong T. Nguyen ◽  
Jeffrey Y. Lai ◽  
Allen T. Lee ◽  
Jens T. Kaiser ◽  
Douglas C. Rees
Keyword(s):  
Abc Transporter ◽  
Conformational Changes ◽  
Abc Transporters ◽  
Binding Protein ◽  
Substrate Binding ◽  
Substrate Uptake ◽  
Transport Activity ◽  
Physiological Concentration ◽  
High Affinity ◽  
Translocation Pathway

TheEscherichia colimethionine ABC transporter MetNI exhibits both high-affinity transport towardl-methionine and broad specificity toward methionine derivatives, includingd-methionine. In this work, we characterize the transport ofd-methionine derivatives by the MetNI transporter. Unexpectedly, the N229A substrate-binding deficient variant of the cognate binding protein MetQ was found to support high MetNI transport activity towardd-selenomethionine. We determined the crystal structure at 2.95 Å resolution of the ATPγS-bound MetNIQ complex in the outward-facing conformation with the N229A apo MetQ variant. This structure revealed conformational changes in MetQ providing substrate access through the binding protein to the transmembrane translocation pathway. MetQ likely mediates uptake of methionine derivatives through two mechanisms: in the methionine-bound form delivering substrate from the periplasm to the transporter (the canonical mechanism) and in the apo form by facilitating ligand binding when complexed to the transporter (the noncanonical mechanism). This dual role for substrate-binding proteins is proposed to provide a kinetic strategy for ABC transporters to transport both high- and low-affinity substrates present in a physiological concentration range.

scholarly journals Biochemical adaptations of two sugar kinases from the hyperthermophilic archaeon Pyrococcus furiosus

2002 ◽  
Vol 366 (1) ◽  
pp. 121-127 ◽  
Author(s):  
Corné H. VERHEES ◽  
Denise G.M. KOOT ◽  
Thijs J.G. ETTEMA ◽  
Cor DIJKEMA ◽  
Willem M. de VOS ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biochemical Characterization ◽  
Pyrococcus Furiosus ◽  
Phosphoryl Group ◽  
Hyperthermophilic Archaeon ◽  
High Affinity ◽  
Optimal Activity ◽  
Genes Encoding ◽  
Modified Embden Meyerhof Pathway

The hyperthermophilic archaeon Pyrococcus furiosus possesses a modified Embden—Meyerhof pathway, including an unusual ADP-dependent glucokinase (ADP-GLK) and an ADP-dependent phosphofructokinase. In the present study, we report the characterization of a P. furiosus galactokinase (GALK) and its comparison with the P. furiosus ADP-GLK. The pyrococcal genes encoding the ADP-GLK and GALK were functionally expressed in Escherichia coli, and the proteins were subsequently purified to homogeneity. Both enzymes are specific kinases with an optimal activity at approx. 90°C. Biochemical characterization of these enzymes confirmed that the ADP-GLK is unable to use ATP as the phosphoryl group donor, but revealed that GALK is ATP-dependent and has an extremely high affinity for ATP. There is a discussion about whether the unusual features of these two classes of kinases might reflect adaptations to a relatively low intracellular ATP concentration in the hyperthermophilic archaeon P. furiosus.

scholarly journals Periplasmic binding protein dependent transport system for maltose and maltodextrins: some recent studies

1989 ◽  
Vol 63 (1-2) ◽  
pp. 53-60
Author(s):  
W. Saurin ◽  
E. Francoz ◽  
P. Martineau ◽  
A. Charbit ◽  
E. Dassa ◽  
...  
Keyword(s):  
Transport System ◽  
Binding Protein ◽  
Periplasmic Binding Protein ◽  
Dependent Transport

scholarly journals The Maltodextrin System of Escherichia coli: Metabolism and Transport

2005 ◽  
Vol 187 (24) ◽  
pp. 8322-8331 ◽  
Author(s):  
Renate Dippel ◽  
Winfried Boos
Keyword(s):  
Escherichia Coli ◽  
Abc Transporter ◽  
Binding Protein ◽  
Glycogen Synthesis ◽  
Equilibrium Conditions ◽  
Per Se ◽  
Maltodextrin Phosphorylase ◽  
Internal Level

ABSTRACT The maltose/maltodextrin regulon of Escherichia coli consists of 10 genes which encode a binding protein-dependent ABC transporter and four enzymes acting on maltodextrins. All mal genes are controlled by MalT, a transcriptional activator that is exclusively activated by maltotriose. By the action of amylomaltase, we prepared uniformly labeled [14C]maltodextrins from maltose up to maltoheptaose with identical specific radioactivities with respect to their glucosyl residues, which made it possible to quantitatively follow the rate of transport for each maltodextrin. Isogenic malQ mutants lacking maltodextrin phosphorylase (MalP) or maltodextrin glucosidase (MalZ) or both were constructed. The resulting in vivo pattern of maltodextrin metabolism was determined by analyzing accumulated [14C]maltodextrins. MalP− MalZ+ strains degraded all dextrins to maltose, whereas MalP+ MalZ− strains degraded them to maltotriose. The labeled dextrins were used to measure the rate of transport in the absence of cytoplasmic metabolism. Irrespective of the length of the dextrin, the rates of transport at a submicromolar concentration were similar for the maltodextrins when the rate was calculated per glucosyl residue, suggesting a novel mode for substrate translocation. Strains lacking MalQ and maltose transacetylase were tested for their ability to accumulate maltose. At 1.8 nM external maltose, the ratio of internal to external maltose concentration under equilibrium conditions reached 106 to 1 but declined at higher external maltose concentrations. The maximal internal level of maltose at increasing external maltose concentrations was around 100 mM. A strain lacking malQ, malP, and malZ as well as glycogen synthesis and in which maltodextrins are not chemically altered could be induced by external maltose as well as by all other maltodextrins, demonstrating the role of transport per se for induction.

scholarly journals High-affinity maltose/trehalose transport system in the hyperthermophilic archaeon Thermococcus litoralis.

1996 ◽  
Vol 178 (16) ◽  
pp. 4773-4777 ◽  
Author(s):  
K B Xavier ◽  
L O Martins ◽  
R Peist ◽  
M Kossmann ◽  
W Boos ◽  
...  
Keyword(s):  
Transport System ◽  
Thermococcus Litoralis ◽  
Hyperthermophilic Archaeon ◽  
High Affinity
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