Structural insights into choline-O-sulfatase reveal the molecular determinants for ligand binding
| Autor: | Jose Antonio Gavira, Ana Cámara-Artigas, Jose Luis Neira, Jesús M. Torres de Pinedo, Pilar Sánchez, Esperanza Ortega, Sergio Martinez-Rodríguez |
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| Přispěvatelé: | Ministerio de Ciencia e Innovación (España), Junta de Andalucía, Universidad de Granada |
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Zaguán. Repositorio Digital de la Universidad de Zaragoza instname Digibug. Repositorio Institucional de la Universidad de Granada |
| Popis: | This work was supported by the Spanish Ministry of Science and Innovation/FEDER grants PID2020-116261GB-I00 (JAG) and RTI2018-097991-B-I00 (JLN), Secretaria General de Universidades, Investigacion y Tecnologia, Junta de Andalucia (PY20-00149 and UAL18-BIO-B005-B; ACA) and the University of Granada (grant PPJI2017-1; SMR). Funding for open access charge: Universidad de Granada/CBUA. Choline-O-sulfatase (COSe; EC 3.1.6.6) is a member of the alkaline phosphatase (AP) superfamily, and its natural function is to hydrolyze choline-O-sulfate into choline and sulfate. Despite its natural function, the major interest in this enzyme resides in the landmark catalytic/substrate promiscuity of sulfatases, which has led to attention in the biotechnological field due to their potential in protein engineering. In this work, an in-depth structural analysis of wild-type Sinorhizobium (Ensifer) meliloti COSe (SmeCOSe) and its C54S active-site mutant is reported. The binding mode of this AP superfamily member to both products of the reaction (sulfate and choline) and to a substrate-like compound are shown for the first time. The structures further confirm the importance of the C-terminal extension of the enzyme in becoming part of the active site and participating in enzyme activity through dynamic intra-subunit and inter-subunit hydrogen bonds (Asn146A–Asp500B–Asn498B). These residues act as the ‘gatekeeper’ responsible for the open/closed conformations of the enzyme, in addition to assisting in ligand binding through the rearrangement of Leu499 (with a movement of approximately 5 A ° ). Trp129 and His145 clamp the quaternary ammonium moiety of choline and also connect the catalytic cleft to the C-terminus of an adjacent protomer. The structural information reported here contrasts with the proposed role of conformational dynamics in promoting the enzymatic catalytic proficiency of an enzyme. Spanish Government European Commission PID2020-116261GB-I00 RTI2018-097991-B-I00 Secretaria General de Universidades Junta de Andalucia PY20-00149 UAL18-BIO-B005-B University of Granada PPJI2017-1 |
| Databáze: | OpenAIRE |
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