PDPK1, Active | 7706

The initial steps in insulin signal transduction occur at the plasma membrane and lead to the activation of phosphatidylinositide (PtdIns) 3-kinase and the formation of PtdIns(3,4,5,)P3 in the inner leaflet of the plasma membrane which is then converted to PtdIns(3,4)P2 by a specific phosphatase. PDK1 or 3-phosphoinositide-dependent protein kinase (PDK1), is activated by the presence of PtdIns(3,4,5)P3 or PtdIns(3,4)P2 (1). PDK1 then activates protein kinase B (PKB) (2) which, in turn, inactivates glycogen synthase kinase-3 (GSK3). The phosphorylation of other proteins by PKB and GSK3 is likely to mediate many of the intracellular actions of insulin. Thus, PDK1 plays a key role in mediating many of the actions of the second messenger(s) PtdIns(3,4, 5)P3 and/or PtdIns(3,4)P2. The human PDK1 is a 556-residue monomeric enzyme comprising of a catalytic domain that is most similar to the PKA, PKB and PKC subfamily of protein kinases and a carboxy-terminal pleckstrin homology (PH) domain. The PDK1 gene is located on human chromosome 16p13.3 and is expressed ubiquitously in human tissues. Human PDK1 is homologous to the Drosophila protein kinase DSTPK61 (3), which have been implicated in the regulation of sex differentiation, oogenesis and spermatogenesis. Expressed PDK1 and DSTPK61 phosphorylated Thr308 of PKB only in the presence of Ptdlns(3,4,5)P3 or Ptdlns(3,4)P2. Overexpression of PDK1 in 293 cells activated PKB and potentiated the IGF1-induced phosphorylation of PKB at Thr308.

7706 | PDK1 Active DataSheet

Biomolecule/Target: N/A

Synonyms: 3-phosphoinositide dependent protein kinase 1; PDK1

Alternates names: 3-phosphoinositide dependent protein kinase 1; PDK1

Taglines: Plays a central role in insulin-mediated signal transduction

NCBI Gene ID #: 6358

NCBI Gene Symbol: CCL14

Gene Source: N/A

Accession #: Q16627

Recombinant: Yes

Source: Baculovirus (Sf9 insect cells)

Purity by SDS-PAGEs: 95%

Assay: SDS-PAGE

Purity: N/A

Assay #2: HPLC

Endotoxin Level: N/A

Activity (Specifications/test method): 159 nmol phosphate incorporated into PDKtide substrate peptide per minute per mg protein at 30°C for 15 minutes using a final concentration of 50 µM ATP (0.83 µCi/assay).

Biological activity: N/A

Results: N/A

Binding Capacity: N/A

Unit Definition: N/A

Molecular Weight: ~67 kDa

Concentration: 5µg/50 µl

Appearance: Liquid

Physical form description: Recombinant proteins in storage buffer (50 mM sodium phosphate, pH 7.0, 300 mM NaCl, 150 mM imidazole, 0.1 mM PMSF, 0.25 mM DTT, 25% glycerol).

Reconstitution Instructions: Dissolve in 1x PBS (It is not recommended to reconstitute to a final concentration less than 100 g/ml.). This can further be diluted to other aqueous buffers.

Amino acid sequence: Target protein is expressed with sequence (Thr20-Asn93) of Human hcc-1 (Uniprot #Q16627) fused with a polyhistidine tag at the C-terminus.