title: Purification, crystallization and preliminary X-ray diffraction of a proteolytic fragment of PDK1 containing the pleckstrin homology domain.
abstract: 3-Phosphoinositide-dependent protein kinase-1 (PDK1) is a Ser/Thr kinase with an essential role in insulin and growth-factor signalling. PDK1 activity towards protein kinase B (PKB) is partially regulated by its pleckstrin homology (PH) domain, which preferentially binds to 3-phosphoinositides. However, the precise molecular mechanism of this regulation is not well understood. Here, the cloning, purification and crystallization of a 150-amino-acid C-terminal region of PDK1 containing the PH domain is reported. A crystal of the PDK1 PH domain grown in the presence of inositol 1,3,4,5-tetrakisphosphate and derivatized with AuCN diffracted to 1.5 A at a synchrotron source. Diffraction data collected near the Au edge resulted in an anomalous Patterson map with a 30sigma peak.
authors.affiliation: Division of Biological Chemistry and Molecular Microbiology, School of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland.
authors.forename: David
authors.initials: D
authors.lastname: Komander
authors.forename: Maria
authors.initials: M
authors.lastname: Deak
authors.forename: Nick
authors.initials: N
authors.lastname: Morrice
authors.forename: Daan M F
authors.initials: DM
authors.lastname: van Aalten
mesh_terms: D064413:3-Phosphoinositide-Dependent Protein Kinases; D001798:Blood Proteins / Q000737:chemistry; D003001:Cloning, Molecular; D003460:Crystallization; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D010750:Phosphoproteins / Q000737:chemistry; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D014961:X-Ray Diffraction
chemical_list: D001798:Blood Proteins; D018076:DNA, Complementary; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C039077:platelet protein P47; D005982:Glutathione Transferase; D064413:3-Phosphoinositide-Dependent Protein Kinases; C578562:PDPK1 protein, human; D017346:Protein Serine-Threonine Kinases