@misc{10481/74975, year = {2022}, month = {4}, url = {http://hdl.handle.net/10481/74975}, abstract = {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.}, organization = {Spanish Government European Commission PID2020-116261GB-I00 RTI2018-097991-B-I00}, organization = {Secretaria General de Universidades}, organization = {Junta de Andalucia PY20-00149 UAL18-BIO-B005-B}, organization = {University of Granada PPJI2017-1}, publisher = {International Union of Crystallography}, keywords = {Choline}, keywords = {Sulfatases}, keywords = {Conformational gating}, keywords = {Alkaline phosphatases}, keywords = {Promiscuity}, title = {Structural insights into choline-O-sulfatase reveal the molecular determinants for ligand binding}, doi = {10.1107/S2059798322003709}, author = {Gavira Gallardo, José Antonio and Torres De Pinedo, Jesús Manuel and Sánchez Medina, María Pilar and Ortega Sánchez, Esperanza and Martínez Rodríguez, Sergio}, }