title: Tris(hydroxymethyl)aminomethane induces conformational change and crystal-packing contraction of porcine pancreatic elastase.
abstract: Porcine pancreatic elastase (PPE) was crystallized under new sulfate-free conditions containing 0.3 M NaCl and 50 mM tris(hydroxymethyl)aminomethane-HCl at pH 7.0. The crystal structure determined at 1.5 angstroms resolution had a unique conformation in four regions which contained loop portions. A chloride ion was bound near the catalytic triad instead of the sulfate ion in PDB entry 1qnj, a typical PPE crystal structure. However, the chloride ion did not affect the configuration of the catalytic triad. A tris(hydroxymethyl)aminomethane (Tris) molecule was bound to the S4 and S5 subsites in place of the adjacent molecule in the 1qnj crystal and played a significant role in the structural change of the region. The distortion in this region may subsequently have induced conformational changes in the other three regions. The fact that Tris and these four regions make a diagonal line in the ac plane may have affected the crystal-packing contraction along the a and c axes in the crystal compared with the typical crystal.
authors.affiliation: Graduate School of Science, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan. kinotk@b.s.osakafu-u.ac.jp
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Kinoshita
authors.forename: Asako
authors.initials: A
authors.lastname: Yamaguchi
authors.forename: Toshiji
authors.initials: T
authors.lastname: Tada
mesh_terms: D000179:Acrylates / Q000494:pharmacology*; D000818:Animals; D001665:Binding Sites; D020134:Catalytic Domain; D002712:Chlorides / Q000494:pharmacology; D018360:Crystallography, X-Ray; D008744:Methylamines / Q000494:pharmacology*; D010196:Pancreatic Elastase / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D013552:Swine
chemical_list: D000179:Acrylates; D002712:Chlorides; D008744:Methylamines; C053576:N-acryloyl-tris(hydroxymethyl)aminomethane; D010196:Pancreatic Elastase