title: Preliminary X-ray diffraction studies of recombinant 19 kDa human fibroblast collagenase.
abstract: Crystals of the catalytic domain of human fibroblast collagenase have been grown in the presence and absence of an inhibitor. Crystals of the inhibitor complex grew from 0.2 M ammonium sulfate and 15 to 30% PEG 8000 at 22 degrees C as bipyramids in the space group P6(2) or P6(4). Crystals of the unligated enzyme grew as rods in the space group P4(1)2(1)2 or P4(3)2(1)2 from 1.0 to 2.0 M sodium formate at 4 degrees C. Both crystal forms grew quite slowly over a period of months, but ultimately yielded crystals that diffracted beyond 2.5 A. The collagenase samples used in these studies were heterogeneous at the amino terminus. Three major species (full length, N-1 and N-2) were identified by mass spectrometry and Edman sequencing. Analysis of dissolved crystals revealed the native crystal form selectively crystallized as the N-2 species; however, no selectivity of N-terminal forms was observed for crystals of the inhibitor complex.
authors.affiliation: Glaxo Research Institute, Department of Bioanalytical, Research Triangle Park, NC 27709.
authors.forename: A M
authors.initials: AM
authors.lastname: Hassell
authors.forename: R J
authors.initials: RJ
authors.lastname: Anderegg
authors.forename: D
authors.initials: D
authors.lastname: Weigl
authors.forename: M V
authors.initials: MV
authors.lastname: Milburn
authors.forename: W
authors.initials: W
authors.lastname: Burkhart
authors.forename: G F
authors.initials: GF
authors.lastname: Smith
authors.forename: P
authors.initials: P
authors.lastname: Graber
authors.forename: T N
authors.initials: TN
authors.lastname: Wells
authors.forename: M A
authors.initials: MA
authors.lastname: Luther
authors.forename: S R
authors.initials: SR
authors.lastname: Jordan
mesh_terms: D017364:Collagenases / Q000648:ultrastructure*; D018360:Crystallography, X-Ray; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D046911:Macromolecular Substances; D013058:Mass Spectrometry; D061965:Matrix Metalloproteinase Inhibitors; D008970:Molecular Weight; D011994:Recombinant Proteins
chemical_list: D046911:Macromolecular Substances; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins; D017364:Collagenases