Recombinant Human CDC27 Protein: Biology & Research Guide

Recombinant Human CDC27 Protein is a key scaffold subunit of the Anaphase-Promoting Complex/Cyclosome (APC/C), playing an essential role in orchestrating cell cycle progression through ubiquitin-mediated proteolysis of mitotic regulators.

Protein Name

Recombinant Human CDC27 Protein

Also Known As

CDC27, APC3, ANAPC3, Cell division cycle 27 homolog

Biological Function

Scaffold subunit of the APC/C E3 ubiquitin ligase complex; recruits co-activators CDC20 and CDH1 to regulate mitotic exit and G1 phase

Structural Feature

Contains multiple tetratricopeptide repeat (TPR) motifs mediating protein-protein interactions within the APC/C

Product SKU

rp365212

Source Reference

absea#PP-776

Species

Human (Homo sapiens)

Biological Background of CDC27

CDC27, also designated APC3 or ANAPC3, is an evolutionarily conserved protein first identified in Saccharomyces cerevisiae as a component required for cell division cycle progression. In humans, the CDC27 gene encodes a large protein characterized by an array of tetratricopeptide repeat (TPR) motifs. These TPR domains serve as molecular scaffolding surfaces that mediate specific protein–protein interactions, making CDC27 an organizational hub within the Anaphase-Promoting Complex/Cyclosome (APC/C).

The APC/C is a multi-subunit E3 ubiquitin ligase that controls the orderly degradation of key cell cycle regulators, including securin and cyclin B, thereby governing the metaphase-to-anaphase transition and mitotic exit. CDC27 acts as a structural scaffold within this complex and is critical for the recruitment of the co-activator proteins CDC20 and CDH1, which confer substrate specificity to the APC/C during different phases of the cell cycle.

Dysregulation of CDC27 has been associated with chromosomal instability and has been identified as a factor in various cancers, making it a subject of interest in both fundamental cell biology and translational oncology research.

Protein Structure and Molecular Characteristics

Human CDC27 is a large protein composed predominantly of TPR (tetratricopeptide repeat) motifs. TPR motifs are degenerate 34-amino-acid repeats that stack together to form a right-handed helical structure, creating a surface well-suited for binding partner proteins. In the context of APC/C, the TPR domains of CDC27 form the primary docking site for the isoleucine–arginine (IR) tail sequences present in the C-termini of co-activators CDC20 and CDH1.

Structural studies of the APC/C, including cryo-electron microscopy data, have revealed that CDC27 occupies a central position on the platform module of the complex, contributing to both the structural integrity of APC/C and the precise positioning of substrate-recognition co-activators. Phosphorylation of CDC27 by mitotic kinases such as CDK1 (cyclin-dependent kinase 1) modulates its interaction with co-activators, providing a regulatory switch that ensures APC/C activity is temporally controlled during the cell cycle.

Recombinant Human CDC27 Protein produced for research purposes retains the characteristic domain architecture necessary for studying these interactions in vitro, enabling biochemical and structural investigations of APC/C assembly and regulation.

Research Applications

Recombinant Human CDC27 Protein is a versatile reagent suitable for multiple research applications in cell cycle biology, biochemistry, and oncology:

• Protein–Protein Interaction Studies: CDC27's TPR domains enable pull-down assays and co-immunoprecipitation experiments to map interactions with APC/C co-activators (CDC20, CDH1) and other binding partners.
• In Vitro Ubiquitylation Assays: As a core APC/C subunit, recombinant CDC27 can be incorporated into reconstituted ubiquitylation systems to study APC/C activity and substrate selectivity.
• Phosphorylation Studies: CDC27 is a substrate of mitotic kinases; recombinant protein enables kinase assays examining how phosphorylation events regulate APC/C assembly and activity.
• Structural Biology: Purified recombinant CDC27 is suitable for use in cryo-EM, X-ray crystallography, and NMR studies aimed at resolving the molecular architecture of APC/C subcomplexes.
• Antibody Validation: Recombinant CDC27 serves as a positive control antigen for validating antibodies used in Western blotting, ELISA, and immunofluorescence assays.
• Drug Discovery: CDC27 and the APC/C pathway represent potential therapeutic targets in cancer; recombinant protein supports screening campaigns for small-molecule inhibitors of APC/C assembly or activity.

Handling, Storage, and Quality Considerations

To ensure optimal performance and data reproducibility when working with Recombinant Human CDC27 Protein, researchers should adhere to established best practices for recombinant protein handling:

• Storage: Store the protein according to the manufacturer's recommended conditions, typically at −70 °C or −80 °C for long-term storage. Avoid repeated freeze–thaw cycles, which can compromise protein integrity and activity.
• Aliquoting: Upon receipt, divide the stock into single-use aliquots before freezing to minimize freeze–thaw degradation.
• Buffer Compatibility: Verify that the supplied storage buffer is compatible with your downstream application. Dialysis or desalting may be required when switching to assay-specific buffers.
• Protein Concentration Verification: Confirm protein concentration using a validated method (e.g., BCA or Bradford assay) in your laboratory prior to use, as shipping conditions may affect sample integrity.
• Quality Control: Examine the protein by SDS-PAGE and, where applicable, by functional assay to confirm expected molecular weight and biological activity before initiating critical experiments.

Refer to the product datasheet (absea#PP-776) for lot-specific specifications including purity, formulation, and endotoxin levels.

Frequently asked questions

What is the primary function of CDC27 in the cell cycle?

CDC27 (APC3/ANAPC3) is a scaffold subunit of the Anaphase-Promoting Complex/Cyclosome (APC/C), an E3 ubiquitin ligase essential for cell cycle progression. It recruits co-activators CDC20 and CDH1 via its TPR domains, enabling the APC/C to ubiquitinate and target key mitotic substrates such as securin and cyclin B for proteasomal degradation, thereby driving mitotic exit and G1 phase entry.

What research applications is Recombinant Human CDC27 Protein suitable for?

Recombinant Human CDC27 Protein is suitable for protein–protein interaction studies (pull-down and co-IP), in vitro ubiquitylation assays, kinase/phosphorylation assays, structural biology investigations (cryo-EM, crystallography), antibody validation (Western blot, ELISA, immunofluorescence), and drug discovery screening targeting the APC/C pathway.

How should Recombinant Human CDC27 Protein be stored to maintain stability?

For long-term stability, Recombinant Human CDC27 Protein should be stored at −70 °C to −80 °C. Repeated freeze–thaw cycles should be avoided; therefore, the protein should be aliquoted into single-use volumes upon receipt. Always refer to the product-specific datasheet (absea#PP-776) for formulation details and any lot-specific storage recommendations.

How is CDC27 regulated during mitosis?

CDC27 is phosphorylated by mitotic kinases, most notably CDK1 (Cyclin B–CDK1 complex), during mitosis. This phosphorylation modulates the affinity of CDC27's TPR domains for the IR-tail motifs of co-activators CDC20 and CDH1, acting as a molecular switch that temporally controls APC/C activation and ensures orderly progression through mitosis and into G1.

Is CDC27 relevant to cancer biology?

Yes, CDC27 has been implicated in cancer biology due to its central role in controlling mitotic exit and maintaining chromosomal stability. Dysregulation or mutation of CDC27 can result in aberrant APC/C activity, leading to chromosomal instability, a hallmark of many cancers. This makes CDC27 a potential biomarker and therapeutic target in oncology research.

References

1. Peters JM. The anaphase-promoting complex: proteolysis in mitosis and beyond. Mol Cell. 2002.
2. Chang L, Zhang Z, Yang J, McLaughlin SH, Barford D. Molecular architecture and mechanism of the anaphase-promoting complex. Nature. 2015.
3. Kraft C, Herzog F, Gieffers C, Mechtler K, Hagting A, Pines J, Peters JM. Mitotic regulation of the human anaphase-promoting complex by phosphorylation. EMBO J. 2003.
4. Sudakin V, Chan GK, Yen TJ. Checkpoint inhibition of the APC/C in HeLa cells is mediated by a dominant inhibitory form of Emi1. J Cell Biol. 2001.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.