title: Stabilization of porcine pancreatic elastase crystals by glutaraldehyde cross-linking.
abstract: Elastase is a serine protease from the chymotrypsin family of enzymes with the ability to degrade elastin, an important component of connective tissues. Excessive elastin proteolysis leads to a number of pathological diseases. Porcine pancreatic elastase (PPE) is often used for drug development as a model for human leukocyte elastase (HLE), with which it shares high sequence identity. Crystals of PPE were grown overnight using sodium sulfate and sodium acetate at acidic pH. Cross-linking the crystals with glutaraldehyde was needed to resist the soaking procedure with a diethyl N-(methyl)pyridinyl-substituted oxo-β-lactam inhibitor. Crystals of PPE bound to the inhibitor belonged to the orthorhombic space group P2₁2₁2₁, with unit-cell parameters a = 51.0, b = 58.3, c = 74.9 Å, and diffracted to 1.8 Å resolution using an in-house X-ray source.
authors.affiliation: Instituto de Tecnologia Química e Biológica - António Xavier, Universidade Nova de Lisboa, Avenida da República, EAN, 2780-157 Oeiras, Portugal.
authors.forename: Stefan
authors.initials: S
authors.lastname: Hofbauer
authors.affiliation: Instituto de Tecnologia Química e Biológica - António Xavier, Universidade Nova de Lisboa, Avenida da República, EAN, 2780-157 Oeiras, Portugal.
authors.forename: José A
authors.initials: JA
authors.lastname: Brito
authors.affiliation: Research Institute for Medicines (i-Med.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Jalmira
authors.initials: J
authors.lastname: Mulchande
authors.affiliation: Instituto de Tecnologia Química e Biológica - António Xavier, Universidade Nova de Lisboa, Avenida da República, EAN, 2780-157 Oeiras, Portugal.
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Nogly
authors.affiliation: Instituto de Tecnologia Química e Biológica - António Xavier, Universidade Nova de Lisboa, Avenida da República, EAN, 2780-157 Oeiras, Portugal.
authors.forename: Miguel
authors.initials: M
authors.lastname: Pessanha
authors.affiliation: Research Institute for Medicines (i-Med.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Rui
authors.initials: R
authors.lastname: Moreira
authors.affiliation: Instituto de Tecnologia Química e Biológica - António Xavier, Universidade Nova de Lisboa, Avenida da República, EAN, 2780-157 Oeiras, Portugal.
authors.forename: Margarida
authors.initials: M
authors.lastname: Archer
mesh_terms: D000818:Animals; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004795:Enzyme Stability; D005976:Glutaral / Q000494:pharmacology*; D010179:Pancreas / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D034421:Sus scrofa; D065093:beta-Lactamase Inhibitors / Q000737:chemistry / Q000494:pharmacology
chemical_list: D003432:Cross-Linking Reagents; D065093:beta-Lactamase Inhibitors; D010196:Pancreatic Elastase; D005976:Glutaral
keywords: glutaraldehyde cross-linking; porcine pancreatic elastase; sensitive protein crystals; serine protease; soaking of hydrophobic ligand; streak-seeding