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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3-Phosphoinositide-Dependent Protein Kinases
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Blood Proteins / chemistry
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Cloning, Molecular
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Crystallization
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DNA, Complementary / metabolism
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Electrophoresis, Polyacrylamide Gel
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Glutathione Transferase / metabolism
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Humans
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Phosphoproteins / chemistry
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Protein Serine-Threonine Kinases / chemistry*
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / isolation & purification
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Protein Structure, Tertiary
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Recombinant Fusion Proteins / metabolism
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Time Factors
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X-Ray Diffraction
Substances
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Blood Proteins
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DNA, Complementary
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Phosphoproteins
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Recombinant Fusion Proteins
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platelet protein P47
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Glutathione Transferase
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3-Phosphoinositide-Dependent Protein Kinases
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PDPK1 protein, human
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Protein Serine-Threonine Kinases