CDK4 recombinant protein

CDK4, Unactive recombinant protein

Cat. Num. AAA515330

• Gene Names: CDK4; CMM3; PSK-J3
• Applications: Western Blot
• Synonyms: CDK4; Unactive recombinant protein; CMM3; MGC14458; PSK-J3; CDK4 recombinant protein

• Host: Sf9 insect cells
• Form/Format: 50mM Tris-HCl, pH 7.5, 150mM NaCl, 10mM glutathione, 0.1mM EDTA, 0.25mM DTT, 0.1mM PMSF, 25% glycerol.
• Sequence Length: 2020

• Applicable Applications for CDK4 recombinant protein: Kinase Assay, Western Blot (WB)
• Type: Recombinant Fusion Protein
• Species: Human
• Tag Information: GST tag
• Expression System: Sf9 insect cells using baculovirus
• Source Note: Recombinant full-length human CDK4 was co-expressed with CDK7 by baculovirus in Sf9 insect cells
• Preparation and Storage: Store product at -70 degree C. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature. For most favorable performance, avoid repeated handling and multiple freeze/thaw cycles.

SDS-PAGE

Related Product Information for CDK4 recombinant protein

Recombinant full-length human CDK4 was co-expressed with CDK7 by baculovirus in Sf9 insect cells using an N-terminal GST tag.

Scientific Background: CDK4 is a member of the cyclin-dependent protein kinases and is involved in the control of cell proliferation during the G1 phase of the cell cycle. CDK4 forms a complex with the D-type cyclins which regulate its activity. CDK4 is inhibited by p16, also known as cyclin-dependent kinase inhibitor-2. CDK4 can mediate phosphorylation of the C-terminal region of Rb protein leading to an active transcriptional repression of E2F complex (1). CDC37 and HSP90 can preferentially associate with the fraction of CDK4 not bound to D-type cyclins. Pharmacologic inactivation of CDC37/HSP90 function leads to reduced stability of CDK4. SMAD3 is a major physiologic substrate of the G1 cyclin-dependent kinases CDK4 and CDK2 (2).

Product Categories/Family for CDK4 recombinant protein

Unactive Kinase; Signaling Proteins - Unactive Kinases

References

1. Harbour, J W. et al: Cdk phosphorylation triggers sequential intramolecular interactions that progressively block Rb functions as cells move through G1. Cell 98: 859-869, 1999.
2. Matsuura, I. et al: Cyclin-dependent kinases regulate the antiproliferative function of Smads. Nature 430: 226-231, 2004.

NCBI and Uniprot Product Information

• NCBI GI #: 345525417
• NCBI GeneID: 1019
• NCBI Accession #: NP_000066.1
• NCBI GenBank Nucleotide #: NM_000075
• UniProt Accession #: P11802; O00576; Q6FG61
• Molecular Weight: ~57 kDa
• NCBI Official Full Name: Homo sapiens cyclin-dependent kinase 4 (CDK4), mRNA
• NCBI Official Synonym Full Names: cyclin-dependent kinase 4
• NCBI Official Symbol: CDK4
• NCBI Official Synonym Symbols: CMM3; PSK-J3
• NCBI Protein Information: cyclin-dependent kinase 4; cell division protein kinase 4
• UniProt Protein Name: Cyclin-dependent kinase 4
• Protein Family: Cyclin-dependent kinase
• UniProt Gene Name: CDK4
• UniProt Entry Name: CDK4_HUMAN

NCBI Description

The protein encoded by this gene is a member of the Ser/Thr protein kinase family. This protein is highly similar to the gene products of S. cerevisiae cdc28 and S. pombe cdc2. It is a catalytic subunit of the protein kinase complex that is important for cell cycle G1 phase progression. The activity of this kinase is restricted to the G1-S phase, which is controlled by the regulatory subunits D-type cyclins and CDK inhibitor p16(INK4a). This kinase was shown to be responsible for the phosphorylation of retinoblastoma gene product (Rb). Mutations in this gene as well as in its related proteins including D-type cyclins, p16(INK4a) and Rb were all found to be associated with tumorigenesis of a variety of cancers. Multiple polyadenylation sites of this gene have been reported. [provided by RefSeq, Jul 2008]

Uniprot Description

Function: Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G1/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G1 phase. Hypophosphorylates RB1 in early G1 phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex. Ref.12 Ref.15 Ref.18

Catalytic activity: ATP + a protein = ADP + a phosphoprotein. Ref.13 Ref.20 Ref.24

Enzyme regulation: Both phosphorylation at Thr-172 and binding of a D-type cyclin are necessary for enzymatic activity. Full activation of the cyclin-D-CDK4 complex appears to require other factors such as recruitment of the substrate via a substrate recruitment motif, and/or formation of the CDKN1B ternary complex. Inhibited by INK4 family members. In resting cells, the non-tyrosine-phosphorylated form of CDKN1B prevents phosphorylation at Thr-172 and inactivation, while, in proliferating cells, tyrosine phosphorylation of CDKN1B allows phosphorylation of Thr-172 of CDK4 and subsequennt activation. Ref.21

Subunit structure: Component of the D-CDK4 complex, composed of CDK4 and some D-type G1 cyclin (CCND1, CCND2 or CCND3). Interacts directly in the complex with CCND1, CCND2 or CCND3. Interacts with SEI1 and ZNF655. Forms a ternary complex, cyclin D-CDK4-CDKN1B, involved in modulating CDK4 enzymatic activity. Interacts directly with CDKN1B (phosphorylated on 'Tyr-88' and 'Tyr-89'); the interaction allows assembly of the cyclin D-CDK4 complex, Thr-172 phosphorylation, nuclear translocation and enhances the cyclin D-CDK4 complex activity. CDK4 activity is either inhibited or enhanced depending on stoichiometry of complex. The non-tyrosine-phosphorylated form of CDKN1B prevents T-loop phosphorylation of CDK4 producing inactive CDK4. Interacts (unphosphorylated form) with CDK2. Also forms ternary complexes with CDKN1A or CDKN2A. Interacts directly with CDKN1A (via its N-terminal); the interaction promotes the assembly of the cyclin D-CDK4 complex, its nuclear translocation and promotes the cyclin D-dependent enzyme activity of CDK4. Ref.13 Ref.14 Ref.16 Ref.17 Ref.18 Ref.20

Subcellular location: Cytoplasm. Nucleus. Membrane. Note: Cytoplasmic when non-complexed. Forms a cyclin D-CDK4 complex in the cytoplasm as cells progress through G1 phase. The complex accumulates on the nuclear membrane and enters the nucleus on transition from G1 to S phase. Also present in nucleoli and heterochromatin lumps. Colocalizes with RB1 after release into the nucleus. Ref.13 Ref.18

Post-translational modification: Phosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B. Ref.17 Ref.20 Ref.21 Ref.24 Ref.25

Involvement in disease: Melanoma, cutaneous malignant 3 (CMM3) [MIM:609048]: A malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.Note: Disease susceptibility is associated with variations affecting the gene represented in this entry. Ref.3 Ref.4 Ref.26 Ref.27

Sequence similarities: Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.Contains 1 protein kinase domain.