Roles of nucleotide substructures in the regulation of cystathionine β-synthase domain-containing pyrophosphatase

Anashkin, Viktor A.; Aksenova, V. A.; Vorobyeva, N. N.; Baykov, Alexander A.

doi:10.1016/j.bbagen.2019.05.010

Cited by 2 publications

(4 citation statements)

References 27 publications

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“…This site accommodated all mono-adenosine phosphates and was likely the pseudosymmetry-related S2 site, “silent” in the wild-type enzyme. Notably, a binding stoichiometry greater than one per Bateman module has also been reported for four CBS2 domain variants of M. thermoacetica CBS-PPase [ 20 ] and authentic CBS domain-containing proteins [ 26 , 27 , 28 , 29 ].…”

Section: Discussionmentioning

confidence: 87%

“…In what follows, we consider the identities of the amino acid residues that control these characteristics (Table 5), speculate on the possible structural mechanisms of this control, and compare the obtained information with that reported for other enzymes regulated via CBS domains. CBS-PPase is differentially regulated by adenosine phosphates-AMP and ADP inhibit it, whereas ATP, cAMP, and Ap4A activate it [9,10,20]. Apart from the polyphosphate chain length, three residues of the CBS1 domain appear to control the direction of the effect: Lys95, Lys100, and Gly118.…”

Section: Discussionmentioning

confidence: 99%

“…CBS-PPase is differentially regulated by adenosine phosphates—AMP and ADP inhibit it, whereas ATP, cAMP, and Ap 4 A activate it [ 9 , 10 , 20 ]. Apart from the polyphosphate chain length, three residues of the CBS1 domain appear to control the direction of the effect: Lys95, Lys100, and Gly118.…”

Section: Discussionmentioning

confidence: 99%

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Specific Mutations Reverse Regulatory Effects of Adenosine Phosphates and Increase Their Binding Stoichiometry in CBS Domain-Containing Pyrophosphatase

Anashkin,

Kirillova,

Orlov

et al. 2024

IJMS

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Regulatory cystathionine β-synthase (CBS) domains are widespread in proteins; however, difficulty in structure determination prevents a comprehensive understanding of the underlying regulation mechanism. Tetrameric microbial inorganic pyrophosphatase containing such domains (CBS-PPase) is allosterically inhibited by AMP and ADP and activated by ATP and cell alarmones diadenosine polyphosphates. Each CBS-PPase subunit contains a pair of CBS domains but binds cooperatively to only one molecule of the mono-adenosine derivatives. We used site-directed mutagenesis of Desulfitobacterium hafniense CBS-PPase to identify the key elements determining the direction of the effect (activation or inhibition) and the “half-of-the-sites” ligand binding stoichiometry. Seven amino acid residues were selected in the CBS1 domain, based on the available X-ray structure of the regulatory domains, and substituted by alanine and other residues. The interaction of 11 CBS-PPase variants with the regulating ligands was characterized by activity measurements and isothermal titration calorimetry. Lys100 replacement reversed the effect of ADP from inhibition to activation, whereas Lys95 and Gly118 replacements made ADP an activator at low concentrations but an inhibitor at high concentrations. Replacement of these residues for alanine increased the stoichiometry of mono-adenosine phosphate binding by twofold. These findings identified several key protein residues and suggested a “two non-interacting pairs of interacting regulatory sites” concept in CBS-PPase regulation.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

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Specific Mutations Reverse Regulatory Effects of Adenosine Phosphates and Increase Their Binding Stoichiometry in CBS Domain-Containing Pyrophosphatase

Anashkin,

Kirillova,

Orlov

et al. 2024

IJMS

Self Cite

View full text Add to dashboard Cite

show abstract

“…CBS-PPase is differentially regulated by adenosine phosphates-AMP and ADP inhibit it, whereas ATP, cAMP, and Ap4A activate it [9,10,18]. Apart from the polyphosphate chain length, three residues of the CBS1 domain appear to control the direction of the effect: Lys95, Lys100, and Gly118.…”

Section: Discussionmentioning

confidence: 99%

Specific Mutations Reverse Regulatory Effects of Adenosine Phosphates and Increase Their Binding Stoichiometry in CBS Domain-Containing Pyrophosphatase

Anashkin,

Kirillova,

Orlov

et al. 2024

Preprint

View full text Add to dashboard Cite

Regulatory cystathionine β-synthase (CBS) domains are widespread in proteins; however, difficulty in structure determination prevents a comprehensive understanding of the underlying regulation mechanism. Tetrameric microbial inorganic pyrophosphatase containing such domains (CBS-PPase) is allosterically inhibited by AMP and ADP and activated by ATP and cell alarmones diadenosine polyphosphates. Each CBS-PPase subunit contains a pair of CBS domains but binds cooperatively only one molecule of the mono-adenosine derivatives. We used site-directed mutagenesis of Desulfitobacterium hafniense CBS-PPase to identify the key elements determining the direction of the effect (activation or inhibition) and the “half-of-the-sites” ligand binding stoichiometry. Seven amino acid residues were selected in CBS1 domain based on the available X-ray structure of the regulatory domains and substituted by alanine and other residues. The interaction of 11 CBS-PPase variants with the regulating ligands was characterized by activity measurements and isothermal titration calorimetry. Lys100 replacement reversed the effect of ADP from inhibition to activation, whereas Lys95 and Gly118 replacements made ADP activator at low concentrations but inhibitor at high concentrations. Replacements of these residues for alanine increased the stoichiometry of mono-adenosine phosphate binding twofold. These findings identify several key protein residues and suggest “two non-interacting pairs of interacting regulatory sites” concept in CBS-PPase regulation.

show abstract

Roles of nucleotide substructures in the regulation of cystathionine β-synthase domain-containing pyrophosphatase

Cited by 2 publications

References 27 publications

Specific Mutations Reverse Regulatory Effects of Adenosine Phosphates and Increase Their Binding Stoichiometry in CBS Domain-Containing Pyrophosphatase

Specific Mutations Reverse Regulatory Effects of Adenosine Phosphates and Increase Their Binding Stoichiometry in CBS Domain-Containing Pyrophosphatase

Specific Mutations Reverse Regulatory Effects of Adenosine Phosphates and Increase Their Binding Stoichiometry in CBS Domain-Containing Pyrophosphatase

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