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PMID: 12169624 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

High resolution crystal structure of the human PDK1 catalytic domain defines the regulatory phosphopeptide docking site.

The EMBO journal ·Vol. 21 ·No. 16 ·2002-08-15 ·Pages 4219-28

Biondi RM, Komander D, Thomas CC, Lizcano JM, Deak M, Alessi DR, van Aalten DM

Abstract

3-phosphoinositide dependent protein kinase-1 (PDK1) plays a key role in regulating signalling pathways by activating AGC kinases such as PKB/Akt and S6K. Here we describe the 2.0 A crystal structure of the PDK1 kinase domain in complex with ATP. The structure defines the hydrophobic pocket termed the "PIF-pocket", which plays a key role in mediating the interaction and phosphorylation of certain substrates such as S6K1. Phosphorylation of S6K1 at its C-terminal PIF-pocket-interacting motif promotes the binding of S6K1 with PDK1. In the PDK1 structure, this pocket is occupied by a crystallographic contact with another molecule of PDK1. Interestingly, close to the PIF-pocket in PDK1, there is an ordered sulfate ion, interacting tightly with four surrounding side chains. The roles of these residues were investigated through a combination of site-directed mutagenesis and kinetic studies, the results of which confirm that this region of PDK1 represents a phosphate-dependent docking site. We discuss the possibility that an analogous phosphate-binding regulatory motif may participate in the activation of other AGC kinases. Furthermore, the structure of PDK1 provides a scaffold for the design of specific PDK1 inhibitors.

MeSH Terms
3-Phosphoinositide-Dependent Protein Kinases Adenosine Triphosphate/metabolism Allosteric Site Amino Acid Sequence Catalytic Domain Crystallography, X-Ray Humans Models, Molecular Molecular Sequence Data Phosphopeptides/metabolism Protein Binding Protein Serine-Threonine Kinases/chemistry,metabolism Sequence Alignment
Chemicals
Phosphopeptides Adenosine Triphosphate 3-Phosphoinositide-Dependent Protein Kinases PDPK1 protein, human Protein Serine-Threonine Kinases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Biondi Ricardo M
Division of Signal Transduction Therapy, School of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
Komander David
Thomas Christine C
Lizcano Jose M
Deak Maria
Alessi Dario R
van Aalten Daan M F
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdf437
Published
2002-08-15
Pages
4219-28
Language
English
Region
England
NLM ID
8208664
PMCID
PMC126174
Subset
IM
Grants
Wellcome Trust · United Kingdom
Databases
PDB
1H1W
External Links
PubMed source DOI: 10.1093/emboj/cdf437 PMC Full Text View journal details
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