PAK4, Active Human, recombinant, expressed in E. coli

Product Number P 7248 Storage Temperature: -70 °C

Synonym: p21-activated Kinase 4 The product is active recombinant, full-length human PAK4 containing an N-terminal GST tag. It is supplied Product Description at a concentration of approximately 100 mg/mL in The p21-activated kinases (PAKs) are serine/threonine 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.25 mM DTT, protein kinases that bind to and, in some cases, are 0.1 mM EGTA and 30% glycerol. stimulated by, activated forms of the small GTPases, Cdc42 and Rac. PAKs are now categorized into two Purity: ³ 80% (SDS-PAGE) subgroups: Group I (PAK1, PAK2, PAK3) is involved in the regulation of cellular processes such as Molecular weight: ~90 kDa transcription, cell morphology, motility, and apoptosis, and Group II (PAK4, PAK5, PAK6) is involved in cell 1,2 Specific Activity: ³ 50 units/mg protein (Bradford). morphology, motility and transformation. Please refer to the Certificate of Analysis for the lot-specific activity. The 591-amino acid PAK4 protein belongs to the Group II PAKs. PAK4 differs from other members of the PAK Unit Definition: One unit will incorporate one nanomole family both in sequence and function. PAK4 transcript of phosphate into the synthetic peptide substrate is expressed in all tissues tested, with highest CKRPRAASFAE per minute at 30 °C at pH 7.2 using expression in prostate, testis, and colon. PAK4 a final concentration of 50 µM [32P] ATP. phosphorylates. autophosphorylates and also activates the JNK pathway. The reorganization of the actin Precautions and Disclaimer cytoskeleton is dependent on PAK4 kinase activity and This product is for R&D use only, not for drug, on its interaction with Cdc42Hs. Unlike other members household, or other uses. Please consult the Material of the PAK family, PAK4 provides a novel link between Safety Data Sheet for information regarding hazards Cdc42Hs and the actin cytoskeleton. The cellular and safe handling practices. locations of PAK4 and Cdc42Hs suggest a role for the 3 Golgi in cell morphogenesis. PAK4 also regulates the Preparation instructions activity of the protein kinase LIM kinase 1 (LIMK1), and For maximum product recovery, after thawing, was shown to interact specifically with LIMK1 in binding centrifuge the vial before removing the cap assays. Immune complex kinase assays revealed that both wild-type and constitutively active PAK4 Storage/Stability phosphorylated LIMK1 even more strongly than PAK1, Stable for at least 12 months when stored as undiluted and activated PAK4 stimulated LIMK1's ability to 4 stock at -70 °C. After initial thawing, store in smaller, phosphorylate cofilin. PAK4 can protect cells from working aliquots at -70 °C. Use the working aliquots apoptosis in response to several different types of immediately upon thawing. Avoid repeated freeze-thaw stimuli by phosphorylating and thereby inhibiting the cycles to prevent denaturing of the protein. Do not proapoptotic protein Bad. This leads to the inhibition of store in a frost-free freezer. effector caspases such as caspase 3. PAK4 can inhibit the death signal early in the caspase cascade, References: antagonizing the activation of initiator caspase 8. This 1. Jaffer, Z.M.and Chernoff, J., p21-activated kinases: role in regulating initiator caspases is an entirely novel three more join the Pak., Int. J. Biochem. Cell Biol., role for the PAK proteins and suggests a new 34, 713-717 (2002). mechanism by which these proteins promote cell 5 2. Bagrodia, S.and Cerione, R.A., Pak to the future., survival. Trends Cell Biol. 9, 350-355 (1999). 3. Abo, A., et al., PAK4, a novel effector for Cdc42Hs, 5. Gnesutta, N.and Minden, A., Death receptor- is implicated in the reorganization of the actin induced activation of initiator caspase 8 is cytoskeleton and in the formation of filopodia., antagonized by serine/threonine kinase PAK4., EMBO J. 17, 6527-6540 (1998). Mol. Cell Biol. 23, 7838-7848 (2003). 4. Dan, C., et al., Cytoskeletal changes regulated by the PAK4 serine/threonine kinase are mediated by AH,PHC 05/05-1 LIM kinase 1 and cofilin., J. Biol. Chem., 276, 32115-32121 (2001).

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