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Tel: USA-760-579-0393 Toll free: 888-462-4959 Fax: USA-760-579-0394 [email protected] http://globozymes.com PRODUCT DATASHEET PAK1 PAK1 is a 65- to 68-kDa Group I member of the protein serine/threonine kinase family whose activity is stimulated by the binding of active Rac and Cdc42 Rho family small GTPases. The PAK enzymes are involved in regulating cell morphology and cytoskeleton organization, cell motility, apoptosis, neuronal functions, cell cycle progression, viral replication and numerous other processes. PAK1 has a C-terminus kinase domain and N-terminus Rac- and Cdc42-binding domain. Binding of Rac and Cdc42 (GLO128-005, GLO128-025, GLO128-050) results in the autophosphorylation and concomitant activation of the kinase, which is then thought to dissociate from the G-proteins to act on downstream targets. Such targets include p38 MAP kinase, JNK, Raf1 and Synapsin I. PAK’s have also been found to interact with protein phosphatase 2A (GLO130-GLO132), ribosomal protein S6 kinase, mixed lineage kinase (MLK2), the SH3SH2 adapter Nck and many other regulatory proteins. The catalytic domains of PAK family members are produced in apoptotic cells via a caspase-dependent pathway. Unlike the native enzymes, the catalytic domains are unaffected by Rac and Cdc42. They undergo rapid autophosphorylation and marked activation. Catalog GLO110-001 Cost/vial $145.20 Source Recombinant human produced GST-tagged in E. coli Purity > 90% by SDS-PAGE, apparent Mr ~ 100-kDa Supplied 1 g per vial in 50 l 50 mM Tris-HCl pH 7.0 buffer containing 14 mM -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, GLO126-010) as substrate, measured in the absence of active Cdc42 and Rac. One unit is the amount of PAK1 that incorporates 1 nmol of phosphate into MBP per min at 30 C. Storage Maintain preparations in aliquots at -70 C. Avoid repeated thawing. References: 1. 2. 3. 4. 5. 6. 7. Manser E et al (1994) “A brain serine/threonine protein kinase activated by Cdc42 and Rac1" Nature 367, 40; Creasy CL & Chernoff J (1995) “Cloning and characterization of a member of the MST subfamily of Ste20-like kinases” J Biol Chem 270, 21695; Nikolic M et al (1998) “The p35/Cdk5 kinase is a neuron-specific Rac effector that inhibits Pak1 activity” Nature 395, 194; Sanders LC et al (1999) “Inhibition of myosin light chain kinase by p21-activated kinase” Science 283, 2083; Sells MA et al (1999) “p21-activated kinase 1 (Pak1) regulates cell motility in mammalian fibroblasts” J Cell Biol 145, 837; Zenke FT et al (1999) “Identification of a central phosphorylation site in p21-activated kinase regulating autoinhibition and kinase activity” J Biol Chem 274, 32565; Chaudhary A et al (2000) “Phosphatidylinositol 3-kinase regulates Raf1 through Pak 8. 9. 10. 11. phosphorylation of serine 338" Curr Biol 10, 551; Graves JD et al (2001) “Both phosphorylation and caspase-mediated cleavage contribute to regulation of the Ste20-like protein kinase Mst1 during CD95/Fas-induced apoptosis” J Biol Chem 276, 14909; Sakurada K et al (2002) “Synapsin I is phosphorylated at Ser603 by p21-activated kinases (PAKs) in vitro and in PC12 cells stimulated with bradykinin” J Biol Chem 277, 45473; Wang RA et al (2003) “Essential functions of p21-activated kinase 1 in morphogenesis and differentiation of mammary glands” J Cell Biol 161, 583; Li Z et al (2003) “Directional sensing requires Gbetagamma-mediated PAK1 and PIXalpha-dependent activation of Cdc42" Cell 114, 215 GloboZymes products are for basic science research purposes only. They are not intended for human use. Tel: USA-760-579-0393 Toll free: 866-891-0439 Fax: 760-579-0394 Email: [email protected] http://globozymes.com