{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,13]],"date-time":"2025-05-13T14:00:47Z","timestamp":1747144847988,"version":"3.40.5"},"reference-count":15,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2021,2,17]],"date-time":"2021-02-17T00:00:00Z","timestamp":1613520000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001411","name":"Indian Council of Medical Research","doi-asserted-by":"publisher","award":["ICMR\u2010NIMR\/PB\/2011\/146"],"award-info":[{"award-number":["ICMR\u2010NIMR\/PB\/2011\/146"]}],"id":[{"id":"10.13039\/501100001411","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["FEBS Open Bio"],"published-print":{"date-parts":[[2021,3]]},"abstract":"<jats:p>Substrate specificity of an enzyme is an important characteristic of its mechanism of action. Investigation of the nucleotide specificity of <jats:italic>Plasmodium\u00a0falciparum<\/jats:italic> succinyl\u2010CoA synthetase (SCS; <jats:italic>Pf<\/jats:italic>SCS) would provide crucial insights of its substrate recognition. Charged gatekeeper residues have been shown to alter the substrate specificity via electrostatic interactions with approaching substrates. The enzyme kinetics of recombinant <jats:italic>Pf<\/jats:italic>SCS (wild\u2010type), generated by refolding of the individual <jats:italic>P.\u00a0falciparum<\/jats:italic> SCS\u03b2 and <jats:italic>Blastocystis<\/jats:italic> SCS\u03b1 subunits, demonstrated ADP\u2010forming activity (<jats:italic>K<\/jats:italic><jats:sub>mATP<\/jats:sub>\u00a0=\u00a048\u00a0\u00b5<jats:sc>m<\/jats:sc>). Further, the introduction of charged gatekeeper residues, either positive (Lys and Lys) or negative (Glu and Asp), resulted in significant reductions in the ATP affinity of <jats:italic>Pf<\/jats:italic>SCS. It is interesting to note that the recombinant <jats:italic>Pf<\/jats:italic>SCS\u03b2 subunit can be refolded to a functional enzyme conformation using <jats:italic>Blastocystis<\/jats:italic> SCS\u03b1, indicating the possibility of subunits swapping among different organisms. These results concluded that electrostatic interactions at the gatekeeper region alone are insufficient to alter the substrate specificity of <jats:italic>Pf<\/jats:italic>SCS, and further structural analysis with a particular focus on binding site architecture is required.<\/jats:p>","DOI":"10.1002\/2211-5463.13034","type":"journal-article","created":{"date-parts":[[2020,11,11]],"date-time":"2020-11-11T08:15:09Z","timestamp":1605082509000},"page":"578-587","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["The nucleotide specificity of succinyl\u2010CoA synthetase of <i>Plasmodium falciparum<\/i> is not determined by charged gatekeeper residues alone"],"prefix":"10.1002","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0550-0162","authenticated-orcid":false,"given":"Kapil","family":"Vashisht","sequence":"first","affiliation":[{"name":"Protein Biochemistry and Engineering Lab, Parasite\u2010Host Biology Group ICMR\u2013National Institute of Malaria Research New Delhi India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pallavi","family":"Singh","sequence":"additional","affiliation":[{"name":"Protein Biochemistry and Engineering Lab, Parasite\u2010Host Biology Group ICMR\u2013National Institute of Malaria Research New Delhi India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sonia","family":"Verma","sequence":"additional","affiliation":[{"name":"Protein Biochemistry and Engineering Lab, Parasite\u2010Host Biology Group ICMR\u2013National Institute of Malaria Research New Delhi India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajnikant","family":"Dixit","sequence":"additional","affiliation":[{"name":"Protein Biochemistry and Engineering Lab, Parasite\u2010Host Biology Group ICMR\u2013National Institute of Malaria Research New Delhi 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