Arresting the Catalytic Arginine in Chlorite Dismutases: Impact on Heme Coordination, Thermal Stability, and Catalysis

<strong>Description of the dataset: </strong> <strong>Data type</strong>: spectroscopic measurements (UV-visible, ECD, EPR), DSC measurements, X-ray crystallography datasets, kinetic measurements, computer simulations and data analysis. Files are in <strong>spc</strong>, <strong>par</strong>, <strong>m, csv, dsx </strong>and<strong> mtz</strong> format Information on <strong>origin of the data</strong>: EPR spectroscopic measurements in <strong>spc</strong> and <strong>par</strong> formats EPR spectroscopic simulation and analyses in <strong>m</strong> format UV-vis spectroscopic measurements in <strong>csv</strong> format ECD measurements in <strong>dsx</strong> format Enzyme activity data in <strong>csv</strong> format DSC measurements in <strong>csv</strong> format X-ray data in <strong>mtz</strong> format Are the data <strong>generated</strong> (e.g. by a machine) or <strong>collected</strong> (e.g. by means of a survey)? UV−vis spectra were recorded using a Cary 60 UV−vis spectrophotometer (Agilent) and a model U-3900 spectrophotometer (Hitachi,Mannheim, Germany) Electronic circular dichroism spectroscopy was performed using Chirascan (Applied Photophysics, Leatherhead, U.K.). X-Band CW-EPR experiments were performed on a Bruker ESP300E spectrometer equipped with a liquid helium cryostat (Oxford Inc.) Enzyme activity was measured polarographically following the release of O2 by using a Clark-type oxygen electrode (Oxygraph Plus; Hansatech Instruments, Norfolk, U.K.). Differential scanning calorimetry experiments were performed on a Micro-Cal PEAQ-DSC Automated instrument (Malvern Panalytical Ltd., Malvern, U.K.) equipped with an autosampler for 96-well plates and controlled by the MicroCal PEAQ-DSC software. Crystallization experiments were performed using the sitting drop vapor diffusion method in SWISSCI MRC three-well crystallization plates (Molecular Dimensions, Newmarket, U.K.). Crystallization drops were set up using a mosquito crystallization robot (TTP Labtech). Commercially available crystallization screens were used for further screening. Crystallization plates were stored in a Formulatrix RI-1000 imaging device at 22 °C. Data were collected at 100 K using a PILATUS 6M detector (25 Hz, 450 μm sensor thickness) at beamline P13 operated by EMBL Hamburg at the PETRA III storage ring (DESY, Hamburg, Germany). Data sets were processed with XDS, and symmetry equivalent reflections merged with XDSCONV. Files in <strong>PARACAT_WP3_20210217_01_EPR</strong> folder includes EPR spectroscopic measurements and computer simulations/analyses, original data are in <strong>spc</strong>/<strong>par</strong> formats; files in <strong>m</strong> format were used to process the data. Files in <strong>PARACAT_WP3_20210217_02_UV-vis </strong>folder includes UV-Vis spectroscopic measurements of pH-titration in <strong>csv</strong> format. Files in <strong>PARACAT_WP3_20210217_03_ECD </strong>folder includes ECD measurements of the far UV (180–260 nm) and visible (260–500 nm) area, as well as unfolding curves in <strong>dsx</strong> format. Files in <strong>PARACAT_WP3_20210217_04_activity </strong>folder includes Clark electrode/activity measurements in<strong> csv</strong> format. Files in <strong>PARACAT_WP3_20210217_05_DSC </strong>folder includes DSC measurements in <strong>csv </strong>format. Files in <strong>PARACAT_WP3_20210217_06_Xray </strong>folder includes pre-processed (from the beamline pipeline) data sets in <strong>mtz</strong> format. NB. See the “READ ME” text file in each subfolder for more detailed information on files organization. <strong>Information on</strong>: <strong>Cld</strong>, chlorite dismutase; <strong><em>C</em>Cld</strong>, chlorite dismutase from Cyanothece sp. PCC7425; <strong>CW</strong>, continuous wave; <strong><em>D</em></strong>, tetragonal; <strong>DSC</strong>, differential circular calorimetry; <strong><em>E</em></strong>, rhombic; <strong>ECD</strong>, electronic circular dichroism; <strong>EPR</strong>, electron paramagnetic resonance; <strong>HS</strong>, high-spin; <strong>LS</strong>, low-spin; <strong><em>Nd</em>Cld</strong>, chlorite dismutase from Candidatus “Nitrospira defluvii”; <strong>PDB</strong>, Protein Data Bank; <strong>ZFS</strong>, zero-field splitting. Units of measurement: Concentration:<strong> mM</strong> (millimolar), <strong>µM</strong> (micromolar), <strong>% w/v</strong> (percentage weigth/volume), <strong>mg/mL</strong> (milligrams per milliliter) Volume: <strong>mL</strong> (milliliters), <strong>µL </strong>(microliters) Wavelength:<strong> nm</strong> (nanometers) Temperature:<strong> °C</strong> (Celsius degrees), <strong>K</strong> (Kelvin degrees) Time:<strong> min</strong> (minutes), <strong>h</strong> (hours) Ellipticity: millidegrees Frequency: <strong>GHz</strong> (gigahertz), <strong>kHz</strong> (kilohertz) Power: <strong>mW</strong> (milliwatt)