Purification and Properties of Adenosine Kinase from Dried Brewer’s Yeast

Leibach, Thomas K.; Spiess, Gudrun I.; Neudecker, T.; Peschke, Günter J.; Puchwein, Gerd; Hartmann, Guido

doi:10.1515/bchm2.1971.352.1.328

Cited by 45 publications

(13 citation statements)

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“…Optimal activity of the recombinant yeast AK was obtained at an ATP : Mg molar ratio of 2 : 1 (data not shown), which is consistent with optimal ratios reported for AK from diverse sources (Leibach et al, 1971;Guranowski, 1979;Moffatt et al, 2000). Kinetic analysis of this recombinant enzyme indicates a K m value of 3 µM for A ( Figure 3 and Table 1).…”

Section: Resultssupporting

confidence: 85%

“…Its dependence on the ATP : Mg 2+ ratio and on the presence of inorganic phosphate and other ions has also been reported (Maj et al, 2000(Maj et al, , 2002. Concerning yeasts, a Saccharomyces cerevisiae AK was purified (Leibach et al, 1971). This work indicated the presence of several apparent isoenzyme forms with molecular weights of 37-39 kDa and a rather limited specificity for the nucleoside to be phosphorylated.…”

Section: Introductionmentioning

confidence: 57%

“…AK is broadly distributed since is found in plants, eukaryotic microorganisms and mammalian tissues, where it plays a key role in the regulation of intracellular adenylate and extracellular A levels (Lindberg et al, 1967;Leibach et al, 1971;Guranowski, 1979;Moffatt et al, 2000). The purified enzyme from different sources exhibits a relatively wide substrate specificity as it tolerates modifications in both the sugar and base moieties (Lindberg et al, 1967;Leibach et al, 1971;Kowaluk et al, 1998;Moffatt et al, 2000). Some biochemical properties of AK have been reported, although no agreement has been reached on the order of substrate binding and product release (Henderson et al, 1972;Mimouni et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

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Expression in Escherichia coli of a recombinant adenosine kinase from Saccharomyces cerevisiae: purification, kinetics and substrate analyses

Barrado

Rodríguez

Jiménez

et al. 2003

Yeast

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The Saccharomyces cerevisiae ADO1 gene is known to encode a homologue of eukaryotic adenosine kinases. This gene was expressed in Escherichia coli as a recombinant protein fused to a polyhistidine tag by using the rhamnose-inducible bacterial promoter rhaB. The recombinant protein was purified to apparent homogeneity and its ability to phosphorylate different substrates was evaluated. Adenosine (K m 3 µM) is its primary substrate. In addition, it also phosphorylates, albeit less efficiently, 3 -deoxyadenosine (cordycepin; K m 1.84 mM) and 3 -amino-3 -deoxyadenosine (K m 0.26 mM). Other kinetic properties of the recombinant enzyme have also been determined.

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Section: Resultssupporting

confidence: 85%

Section: Introductionmentioning

confidence: 57%

Section: Introductionmentioning

confidence: 99%

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Expression in Escherichia coli of a recombinant adenosine kinase from Saccharomyces cerevisiae: purification, kinetics and substrate analyses

Barrado

Rodríguez

Jiménez

et al. 2003

Yeast

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“…ADK activity was first characterized from yeast and mammalian tissues (7,33). Subsequently, ADKs have been investigated biochemically and/or genetically from a large number of other eukaryotes, including humans and other mammals (21,44,46,53,70), plants (20,47), moss (66), parasitic protozoa (8,14,16,23), and yeast (36,37). The ADK enzyme has been considered to be the key metabolic regulator for maintenance of certain intra-and extracellular levels of adenosine in eukaryotes (2,22), and there is abundant evidence to indicate that adenosine is a significant signal molecule engaged in regulation of physiology and modulation of the function of various cell types in mammals (6,11,35).…”

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confidence: 99%

An Adenosine Kinase Exists inXanthomonas campestrisPathovar campestris and Is Involved in Extracellular Polysaccharide Production, Cell Motility, and Virulence

Yong-qin

et al. 2009

J Bacteriol

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Adenosine kinase (ADK) is a purine salvage enzyme and a typical housekeeping enzyme in eukaryotes which catalyzes the phosphorylation of adenosine to form AMP. Since prokaryotes synthesize purines de novo and no endogenous ADK activity is detectable in Escherichia coli, ADK has long been considered to be rare in bacteria. To date, only two prokaryotes, both of which are gram-positive bacteria, have been reported to contain ADK. Here we report that the gram-negative bacterium Xanthomonas campestris pathovar campestris, the causal agent of black rot of crucifers, possesses a gene (designated adk Xcc ) encoding an ADK (named ADK Xcc ), and we demonstrate genetically that the ADK Xcc is involved in extracellular polysaccharide (EPS) production, cell motility, and pathogenicity of X. campestris pv. campestris. adk Xcc was overexpressed as a His 6 -tagged protein in E. coli, and the purified His 6 -tagged protein exhibited ADK activity. Mutation of adk Xcc did not affect bacterial growth in rich and minimal media but led to an accumulation of intracellular adenosine and diminutions of intracellular ADK activity and ATP level, as well as EPS. The adk Xcc mutant displayed significant reductions in bacterial growth and virulence in the host plant.

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“…Adenosine kinases have been purified from a number of eukaryotic sources, including human placenta and liver [4,5], rabbit liver [6], yeast [7] and Leishmania donovani [4], and extensively characterized at a kinetic level. Important results have been achieved for the leishmanial enzyme, demonstrating that it displays many specific characteristics compared with other adenosine kinases including that from the natural host of this parasite [4,5,8].…”

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confidence: 99%

Characterization and molecular cloning of an adenosine kinase from Babesia canis rossi

Carret¹,

Delbecq²,

Labesse

et al. 1999

European Journal of Biochemistry

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In the search for immunoprotective antigens of the intraerythrocytic Babesia canis rossi parasite, a new cDNA was cloned and sequenced. Protein sequence database searches suggested that the 41-kDa protein belongs to the phosphofructokinase B type family (PFK-B). However, because of the low level sequence identity (, 20%) of the protein both with adenosine and sugar kinases from this family, its structural and functional features were further investigated using molecular modelling and enzymatic assays. The sequence/structure comparison of the protein with the crystal structure of a member of the PFK-B family, Escherichia coli ribokinase (EcRK), suggested that it might also form a stable and active dimer and revealed conservation of the ATP-binding site. However, residues specifically involved in the ribose-binding sites in the EcRK sequence (S and N) were substituted in its sequence (by H and M, respectively), and were suspected of binding adenosine compounds rather than sugar ones. Enzymatic assays using a purified glutathione S-transferase fusion protein revealed that this protein exhibits rapid catalysis of the phosphorylation of adenosine with an apparent K m value of 70 nm, whereas it was inactive on ribose or other carbohydrates. As enzymatic assays confirmed the results of the structure/function analysis indicating a preferential specificity towards adenosine compounds, this new protein of the PFK-B family corresponds to an adenosine kinase from B. canis rossi. It was named BcrAK.

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Purification and Properties of Adenosine Kinase from Dried Brewer’s Yeast

Cited by 45 publications

References 0 publications

Expression in Escherichia coli of a recombinant adenosine kinase from Saccharomyces cerevisiae: purification, kinetics and substrate analyses

Expression in Escherichia coli of a recombinant adenosine kinase from Saccharomyces cerevisiae: purification, kinetics and substrate analyses

An Adenosine Kinase Exists inXanthomonas campestrisPathovar campestris and Is Involved in Extracellular Polysaccharide Production, Cell Motility, and Virulence

Characterization and molecular cloning of an adenosine kinase from Babesia canis rossi

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