Crystallization and X-ray analysis of the Y75N mutant of Mucor pusillus pepsin complexed with inhibitor

Mohammed O. Badasso, V. Dhanaraj, S. P. Wood, J. B. Cooper, T. L. Blundell

Research output: Contribution to journalArticlepeer-review

Abstract

Y75N mutant Mucor pusillus pepsin has been overexpressed in yeast, purified and cocrystallized with the iodine-containing human renin inhibitor CP-113972 {(2R,3S)-isopropyl 3-[(L-prolyl-p-iodo-L-phenylalanyl-S-methyl-cysteinyl)amino-4]-cyclohexyl-2-hydroxybutanoate} for X-ray crystallography. Tetragonal complex crystals with space group P43212 were produced by the hanging-drop vapour-diffusion method and diffracted to 3.0 Å. The crystals exhibited unit-cell parameters a = b = 182.5, c = 99.1 Å and contained four molecules in the asymmetric unit. A 96% complete data set was collected at 298 K using Cu K[alpha] X-rays from a rotating-anode generator. Solution of the crystal structure of Y75N mutant M. pusillus pepsin is under way by molecular replacement using the molecular coordinates of wild-type M. pusillus pepsin as a model.
Original languageEnglish
Pages (from-to)770-772
JournalActa Crystallographica Section D Biological Crystallography
Volume60
Issue number4
DOIs
Publication statusPublished - 1 Apr 2004

Keywords

  • pepsins
  • aspartic proteinases
  • inhibitors

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