PAK1 Protein Kinase

Source: GST-tagged recombinant human produced in E. coli

Purity: ~ 90% by SDS-PAGE, apparent Mr ~ 100-kDa.

Supplied: In 50 μl 50 mM Tris-HCl pH 7.0 buffer containing 14 mM β-mercaptoethanol, 1 mM benzamidine, 0.1 mM phenylmethanesulfonyl fluoride, 1 mM EDTA, 0.03% Brij-35 and 10% glycerol.

Activity: ~ 200 units/mg with myelin basic protein (MBP) as substrate measured in the absence of Cdc42. One unit is the amount of PAK1 that incorporates 1 nmol of phosphate into MBP per min. Maintain preparations in aliquots at -70° C. Avoid repeated thawing.

Synonyms: GST-PAK1; GST-p65PAK; GST-p68PAK; GST p21-Activated Protein Kinase

Background: PAK1 is a 65-68 kDa Group I member of a protein serine/threonine kinase family whose activity is stimulated by the Rho family small GTPases Rac and Cdc42. PAKs regulate numerous cellular processes. These include cell morphology and cytoskeleton organization, cell motility, apoptosis, neuronal functions, cell cycle progression and viral replication. PAK1 has a C-terminus kinase domain and N-terminus Rac- and Cdc42-binding domain. Binding of Rac and Cdc42 to the N-terminus results in the autophosphorylation and activation of the kinase, which then dissociates from Rac and Cdc42 to act on down stream targets. Such targets include p38 MAP kinase, JNK, cRaf1 and Synapsin I. PAK’s also interact with Protein Phosphatase 2A (PP2A1), ribosomal protein S6 kinase, mixed lineage kinase (MLK2), the SH3-SH2 adapter Nck and other regulatory proteins. The catalytic domains of PAKs are produced in apoptotic cells via a caspase-dependent pathway.  Unlike the native enzymes, do not require Rac and Cdc42 to undergo rapid autophosphorylation and activation.

References: Nature 367: 40; J Biol Chem 270: 21695

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