{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Hu S"],"funding":["Howard Hughes Medical Institute","Office of the President, University of California","National Institute of General Medical Sciences","David and Lucile Packard Foundation","NIGMS NIH HHS","National Science Foundation"],"pagination":["1555-1567"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC6353671"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["141(4)"],"pubmed_abstract":["Soybean lipoxygenase (SLO) has served as a prototype for understanding the molecular origin of enzymatic rate accelerations. The double mutant (DM) L546A/L754A is considered a dramatic outlier, due to the unprecedented size and near temperature-independence of its primary kinetic isotope effect, low catalytic efficiency, and elevated enthalpy of activation. To uncover the physical basis of these features, we herein apply three structural probes: hydrogen-deuterium exchange mass spectrometry, room-temperature X-ray crystallography and EPR spectroscopy on four SLO variants (wild-type (WT) enzyme, DM, and the two parental single mutants, L546A and L754A). DM is found to incorporate features of each parent, with the perturbation at position 546 predominantly influencing thermally activated mot"],"journal":["Journal of the American Chemical Society"],"pubmed_title":["Biophysical Characterization of a Disabled Double Mutant of Soybean Lipoxygenase: The \"Undoing\" of Precise Substrate Positioning Relative to Metal Cofactor and an Identified Dynamical Network."],"pmcid":["PMC6353671"],"funding_grant_id":["GM124149","GM123159","F32 GM113432","R35 GM118117","LFR-17-476732","P30 GM124169","1R35GM126961-01","T32 GM064337","R35 GM126961","1231306","GM118117","R01 GM124149","R01 GM123159"],"pubmed_authors":["Britt RD","Hu S","Carr CAM","Thompson EM","Wilcoxen J","Offenbacher AR","Gee CL","Prigozhin DM","Fraser JS","Klinman JP","Alber T","Yang V"],"additional_accession":[]},"is_claimable":false,"name":"Biophysical Characterization of a Disabled Double Mutant of Soybean Lipoxygenase: The \"Undoing\" of Precise Substrate Positioning Relative to Metal Cofactor and an Identified Dynamical Network.","description":"Soybean lipoxygenase (SLO) has served as a prototype for understanding the molecular origin of enzymatic rate accelerations. The double mutant (DM) L546A/L754A is considered a dramatic outlier, due to the unprecedented size and near temperature-independence of its primary kinetic isotope effect, low catalytic efficiency, and elevated enthalpy of activation. To uncover the physical basis of these features, we herein apply three structural probes: hydrogen-deuterium exchange mass spectrometry, room-temperature X-ray crystallography and EPR spectroscopy on four SLO variants (wild-type (WT) enzyme, DM, and the two parental single mutants, L546A and L754A). DM is found to incorporate features of each parent, with the perturbation at position 546 predominantly influencing thermally activated mot","dates":{"release":"2019-01-01T00:00:00Z","publication":"2019 Jan","modification":"2026-05-07T14:01:18.363Z","creation":"2026-04-07T22:48:24.173Z"},"accession":"S-EPMC6353671","cross_references":{"pubmed":["30645119"],"doi":["10.1021/jacs.8b10992"]}}