Thermococcus litoralis uses a modified Embden-Meyerhof pathway. Its phosphofructokinase (TLPFK) catalyses the phosphorylation of fructose-6-phosphate by ADP, but not by ATP, in the presence of Mg(2+), yielding fructose-1,6-bisphosphate. The gene encoding TLPFK was cloned and overexpressed in Escherichia coli. Recombinant TLPFK consists of a dimer with a 52 kDa subunit. The native crystals belong to space group P4(1)2(1)2, with unit-cell parameters a = b = 85.41, c = 163.93 A, and diffract to beyond 2.6 A resolution. The TLPFK structure was preliminarily analyzed by means of multiple isomorphous replacement with four heavy-atom derivatives.

Download full-text PDF

Source
http://dx.doi.org/10.1107/s0907444903010631DOI Listing

Publication Analysis

Top Keywords

archaeal adp-dependent
4
adp-dependent phosphofructokinase
4
phosphofructokinase expression
4
expression purification
4
purification crystallization
4
crystallization preliminary
4
preliminary crystallographic
4
crystallographic analysis
4
analysis thermococcus
4
thermococcus litoralis
4

Similar Publications

Kinetic characterization and phylogenetic analysis of human ADP-dependent glucokinase reveal new insights into its regulatory properties.

Arch Biochem Biophys

June 2023

Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile. Electronic address:

Although ADP-dependent sugar kinases were first described in archaea, at present, the presence of an ADP-dependent glucokinase (ADP-GK) in mammals is well documented. This enzyme is mainly expressed in hematopoietic lineages and tumor tissues, although its role has remained elusive. Here, we report a detailed kinetic characterization of the human ADP-dependent glucokinase (hADP-GK), addressing the influence of a putative signal peptide for endoplasmic reticulum (ER) destination by characterizing a truncated form.

View Article and Find Full Text PDF

Methanogenic archaea have received attention due to their potential use in biotechnological applications such as methane production, so their metabolism and regulation are topics of special interest. When growing in a nutrient-rich medium, these organisms exhibit gluconeogenic metabolism; however, under starvation conditions, they turn to glycolytic metabolism. To date, no regulatory mechanism has been described for this gluconeogenic/glycolytic metabolic switch.

View Article and Find Full Text PDF

Identification and Enzymatic Analysis of an Archaeal ATP-Dependent Serine Kinase from the Hyperthermophilic Archaeon .

J Bacteriol

July 2021

Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.

Serine kinase catalyzes the phosphorylation of free serine (Ser) to produce -phosphoserine (Sep). An ADP-dependent Ser kinase in the hyperthermophilic archaeon Thermococcus kodakarensis (-SerK) is involved in cysteine (Cys) biosynthesis and most likely Ser assimilation. An ATP-dependent Ser kinase in the mesophilic bacterium Staphylococcus aureus is involved in siderophore biosynthesis.

View Article and Find Full Text PDF

Structure of an ancestral ADP-dependent kinase with fructose-6P reveals key residues for binding, catalysis, and ligand-induced conformational changes.

J Biol Chem

August 2021

Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile. Electronic address:

ADP-dependent kinases were first described in archaea, although their presence has also been reported in bacteria and eukaryotes (human and mouse). This enzyme family comprises three substrate specificities; specific phosphofructokinases (ADP-PFKs), specific glucokinases (ADP-GKs), and bifunctional enzymes (ADP-PFK/GK). Although many structures are available for members of this family, none exhibits fructose-6-phosphate (F6P) at the active site.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!