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Recombinant human CDK4/CycD1, active protein kinase,10 µg  

Recombinant human CDK4/CycD1, active protein kinase,10 µg

Recombinant human Cyclin-dependent kinase 4 in complex with cyclin D1 (CDK4/CycD1), active enzyme

Alternate names: CMM3, MGC14458, CMM3, cyclin dependent kinase 4, cell division kinase 4/Cyclin D1, PRAD1, BCL1, U21B31, BCL-1, CDK4, CycD1

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475,00 €

Cdks are the catalytic subunits of serine/threonine protein kinases which are involved in the progression through the cell cycle. The catalytic activity of these kinases requires interaction with cyclins, which are synthesized and degraded during the cell cycle in a cyclical fashion. Induced by mitogenic signals cyclins of the D-type family (D1, D2, and D3) are synthesized and bind and activate CDK4 and CDK6. Active complexes are involved in entering the G1 phase and regulate the early phase of G1 by phosphorylation of the retinoblastoma protein (Rb), p107 and p130. The cyclin D1 gene is amplified and/or overexpressed in various human cancers. For this reason, cyclin-CDK complexes have been considered as very promising therapeutic targets in human malignancies.

Protein: recombinant human CDK4 (amino acids S4-E303, as in NCBI/Protein entry   NP_000066.1) and human CycD1 (Amino acids Q4-I295, as in NCBI/Protein entry NP_444284.1), contain N-terminal GST fusion tags with Thrombin cleavage sites)
Expression system: Sf9 cells
Purification: One-step affinity purification using glutathione agarose
Storage buffer: 50 mM HEPES pH 7.5, 100 mM NaCl, 5 mM DTT, 4 mM reduced glutathione, 20% glycerol
Protein concentration: 0.314 mg/ml (Bradford method using BSA as standard protein)
Method for determination of Km value & specific activity: Filter binding assay MAFC membrane
Specific activity: 99,000 pmol/mg min
Activation: This kinase was not activated by special procedures

Entrez Gene ID: 1019/595 
UniProtKB:  P11802/P24385

Ordering information: shipped on dry ice

Product specific literature references:

Matsushime H, Ewen ME, Sherr CJ et al. (1992) "Identification and properties of an atypical catalytic subunit (p34PSK-J3/cdk4) for mammalian D type G1 cyclins." Cell 71(2):323-34

Serrano M, Hannon GJ, Beach D (1993) "A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4" Nature 366 (6456):704-7

Kato J, Matsushime H, Hiebert SW, Ewen ME, Sherr CJ (1993) "Direct binding of cyclin D to the retinoblastoma gene product (pRb) and pRb phosphorylation by the cyclin D-dependent kinase CDK4." Genes Dev. 7(3):331-42

Tam SW, Theodoras AM, Shay JW, Draetta GF, Pagano M (1994) "Differential expression and regulation of Cyclin D1 protein in normal and tumor human cells: association with Cdk4 is required for Cyclin D1 function in G1 progression." Oncogene 9(9):2663-74

Terada Y, Tatsuka M, Jinno S, Okayama H (1995) "Requirement for tyrosine phosphorylation of Cdk4 in G1 arrest induced by ultraviolet irradiation." Nature 376(6538):358-62

Lukas J, Bartkova J, Rohde M, Strauss M, Bartek J (1995) "Cyclin D1 is dispensable for G1 control in retinoblastoma gene-deficient cells independently of cdk4 activity." Mol Cell Biol. 15(5):2600-11

Hirai H, Roussel MF, Kato JY, Ashmun RA, Sherr CJ (1995) "Novel INK4 proteins, p19 and p18, are specific inhibitors of the cyclin D-dependent kinases CDK4 and CDK6." Mol Cell Biol. 15(5):2672-81

Zuo L, Weger J, Dracopoli NC et al. (1996) "Germline mutations in the p16INK4a binding domain of CDK4 in familial melanoma." Nat Genet. 12(1):97-9

Kitagawa M, Higashi H, Taya Y et al. (1996) "The consensus motif for phosphorylation by cyclin D1-Cdk4 is different from that for phosphorylation by cyclin A/E-Cdk2."  EMBO J. 15(24):7060-9

Rane SG, Dubus P, Mettus RV, Galbreath EJ, Boden G, Reddy EP, Barbacid M (1999) "Loss of Cdk4 expression causes insulin-deficient diabetes and Cdk4 activation results in beta-islet cell hyperplasia." Nat Genet. 22(1):44-52

Hermeking H, Rago C, Kinzler KW et al. (2000) "Identification of CDK4 as a target of c-MYC." Proc Natl Acad Sci U S A. 97(5):2229-34

Miliani de Marval PL, Macias E, Rodriguez-Puebla ML et al. (2004) "Lack of cyclin-dependent kinase 4 inhibits c-myc tumorigenic activities in epithelial tissues." Mol Cell Biol. 24(17):7538-47