title: Crystallization of PTP Domains.
abstract: Protein crystallography is the most powerful method to obtain atomic resolution information on the three-dimensional structure of proteins. An essential step towards determining the crystallographic structure of a protein is to produce good quality crystals from a concentrated sample of purified protein. These crystals are then used to obtain X-ray diffraction data necessary to determine the 3D structure by direct phasing or molecular replacement if the model of a homologous protein is available. Here, we describe the main approaches and techniques to obtain suitable crystals for X-ray diffraction. We include tools and guidance on how to evaluate and design the protein construct, how to prepare Se-methionine derivatized protein, how to assess the stability and quality of the sample, and how to crystallize and prepare crystals for diffraction experiments. While general strategies for protein crystallization are summarized, specific examples of the application of these strategies to the crystallization of PTP domains are discussed.
authors: Manchester Protein Structure Facility, Manchester Institute of Biotechnology, Manchester, UK.
authors: Colin
authors: C
authors: Levy
authors: Faculty of Life Sciences, University of Manchester, Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK.
authors: James
authors: J
authors: Adams
authors: Faculty of Life Sciences, University of Manchester, Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK. Lydia.Tabernero@manchester.ac.uk.
authors: Lydia
authors: L
authors: Tabernero
mesh_terms: D000818:Animals; D003451:Cryoprotective Agents / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods; D005615:Freezing; D005990:Glycerol / Q000737:chemistry; D006801:Humans; D008715:Methionine / Q000737:chemistry; D009821:Oils / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D000072417:Protein Domains; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry*; D012643:Selenium / Q000737:chemistry
chemical_list: D003451:Cryoprotective Agents; D009821:Oils; D011092:Polyethylene Glycols; D008715:Methionine; D017027:Protein Tyrosine Phosphatases; D012643:Selenium; D005990:Glycerol
keywords: Cryoprotectant; Crystal seeding; Crystallization trials; Crystallogenesis; Se-methionine protein