Computational protocol: Arginine 104 Is a Key Catalytic Residue in Leukotriene C4 Synthase*

Similar protocols

Protocol publication

[…] Crystals of R31Q hLTC4S were grown and cryo-cooled as described (). Diffraction data were collected to a resolution of 2.1 Å for the complex at beamline ID14-1 at the European Synchrotron Radiation Facility using an ADSC Quantum Q210 detector. Each frame was exposed for 1 s with an oscillation range of 0.5°. The XDS package () was used for indexing and integration. Scaling of the data was done using SCALA in the CCP4 suite (). Native hLTC4S (Protein Data Bank code 2uuh) without ligands or water molecules was used for initial rigid body refinement of the R31Q hLTC4S data. The Fo − Fc maps produced showed clear electron density around position 31 corresponding to the mutation of Arg to Gln. Electron density corresponding to a GSH molecule was also seen in the active site. REFMAC5 () in the CCP4i package was used for refinement of coordinates and B-factors. COOT () was used for molecular modeling. [...] The refinement of the R31Q hLTC4S structure converged at Rcryst and Rfree of 18 and 20%, respectively. The quality of the final structure was verified using PROCHECK (). The Ramachandran plot for the R31Q hLTC4S structure showed that 97% of residues were in the most favorable regions and 2% were in the allowed region. The coordinates and structure factors for R31Q hLTC4S have been deposited in the Protein Data Bank (code 3LEO). All figures were made using PyMOL (DeLano Scientific). […]

Pipeline specifications

Software tools XDS, CCP4, REFMAC5, Coot, PROCHECK, PyMOL
Applications Small-angle scattering, Protein structure analysis
Organisms Dipturus trachyderma, Homo sapiens
Diseases Asthma
Chemicals Glutathione