{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["2013"],"submitter":["Preobrazhenskaya YV"],"funding":["Warsaw University, Poland"],"pubmed_abstract":["The transformation of the strain DH5α (TM)-T1(R) with plasmid vector pET11a containing the cloned gene of bacterial selenophosphate synthetase (SPS), selD, from the E. coli BL21-Gold (DE3) strain gives an overproducing strain of SPS with one synonymic substitution, E197D. The transformation efficiency was estimated as 8 × 10(8) CFU/ μ g plasmid DNA. 28 mg of highly purified preparation of recombinant SPS capable of binding TNP-ATP was eluted from DEAE-Sephadex column in amount of 15 % from the total soluble protein in crude extract. The fluorescent derivative of ATP, 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-triphosphate (TNP-ATP), was used as a synthetic analog of the substrate for the monitoring and quantitative analysis of the functional activity of SPS. The non-linear regression anal"],"journal":["Journal of amino acids"],"pagination":["983565"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC3956282"],"repository":["biostudies-literature"],"pubmed_title":["Binding Stoichiometry of a Recombinant Selenophosphate Synthetase with One Synonymic Substitution E197D to a Fluorescent Nucleotide Analog of ATP, TNP-ATP."],"pmcid":["PMC3956282"],"pubmed_authors":["Stenko AI","Lugovtsev VY","Preobrazhenskaya YV","Shvarts MV"],"additional_accession":[]},"is_claimable":false,"name":"Binding Stoichiometry of a Recombinant Selenophosphate Synthetase with One Synonymic Substitution E197D to a Fluorescent Nucleotide Analog of ATP, TNP-ATP.","description":"The transformation of the strain DH5α (TM)-T1(R) with plasmid vector pET11a containing the cloned gene of bacterial selenophosphate synthetase (SPS), selD, from the E. coli BL21-Gold (DE3) strain gives an overproducing strain of SPS with one synonymic substitution, E197D. The transformation efficiency was estimated as 8 × 10(8) CFU/ μ g plasmid DNA. 28 mg of highly purified preparation of recombinant SPS capable of binding TNP-ATP was eluted from DEAE-Sephadex column in amount of 15 % from the total soluble protein in crude extract. The fluorescent derivative of ATP, 2'(3')-O-(2,4,6-trinitrophenyl)adenosine-5'-triphosphate (TNP-ATP), was used as a synthetic analog of the substrate for the monitoring and quantitative analysis of the functional activity of SPS. The non-linear regression anal","dates":{"release":"2013-01-01T00:00:00Z","publication":"2013","modification":"2025-04-04T22:45:50.775Z","creation":"2019-03-27T01:23:31Z"},"accession":"S-EPMC3956282","cross_references":{"pubmed":["24719756"],"doi":["10.1155/2013/983565"]}}