Purification, crystallization and preliminary X-ray diffraction of a proteolytic fragment of PDK1 containing the pleckstrin homology domain

Authors: Komander, David; Deak, Maria; Morrice, Nick; van Aalten, Daan M. F.

Source: Acta Crystallographica Section D, Volume 60, Number 2, 1 February 2004 , pp. 314-316(3)

Publisher: Blackwell Publishing

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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 Å at a synchrotron source. Diffraction data collected near the Au edge resulted in an anomalous Patterson map with a 30σ peak.

Keywords: 3-phosphoinositide-dependent protein kinase-1; pleckstrin homology domains

Document Type: Research article

DOI: 10.1107/S0907444903028518

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