title: Crystal engineering yields crystals of cyclophilin D diffracting to 1.7 A resolution.
abstract: In the pharmaceutical industry, knowledge of the three-dimensional structure of a specific target facilitates the drug-discovery process. Despite possessing favoured analytical properties such as high purity and monodispersion in light scattering, some proteins are not capable of forming crystals suitable for X-ray analysis. Cyclophilin D, an isoform of cyclophilin that is expressed in the mitochondria, was selected as a drug target for the treatment of cardiac disorders. As the wild-type enzyme defied all attempts at crystallization, protein engineering on the enzyme surface was performed. The K133I mutant gave crystals that diffracted to 1.7 A resolution using in-house X-ray facilities and were suitable for soaking experiments. The crystals were very robust and diffraction was maintained after soaking in 25% DMSO solution: excellent conditions for the rapid analysis of complex structures including crystallographic fragment screening.
authors.affiliation: F. Hoffmann-La Roche Ltd, Pharmaceutical Research Discovery, CH-4070 Basel, Switzerland. daniel.schlatter@roche.com
authors.forename: Daniel
authors.initials: D
authors.lastname: Schlatter
authors.forename: Ralf
authors.initials: R
authors.lastname: Thoma
authors.forename: Erich
authors.initials: E
authors.lastname: Küng
authors.forename: Martine
authors.initials: M
authors.lastname: Stihle
authors.forename: Francis
authors.initials: F
authors.lastname: Müller
authors.forename: Edilio
authors.initials: E
authors.lastname: Borroni
authors.forename: Andrea
authors.initials: A
authors.lastname: Cesura
authors.forename: Michael
authors.initials: M
authors.lastname: Hennig
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004121:Dimethyl Sulfoxide; D008027:Light; D016297:Mutagenesis, Site-Directed; D019696:Peptidylprolyl Isomerase / Q000737:chemistry; D010957:Plasmids / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D015202:Protein Engineering; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012542:Scattering, Radiation; D012997:Solvents; D013050:Spectrometry, Fluorescence; D014461:Ultracentrifugation
chemical_list: D000081406:Peptidyl-Prolyl Isomerase F; D012997:Solvents; D004247:DNA; D021983:Cyclophilins; D019696:Peptidylprolyl Isomerase; D004121:Dimethyl Sulfoxide