Advertisement

Expression and Purification of Recombinant Cyclins and CDKs for Activity Evaluation

  • Edurne Gallastegui
  • Oriol BachsEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1336)

Abstract

Cyclin-dependent kinases (Cdks) belong to a family of key regulators of cell division cycle and transcription. Their activity is mainly regulated by association with regulatory subunits named cyclins but their activities are also regulated by phosphorylation, acetylation, and the association with specific inhibitory proteins (CKIs). The activity of different Cdks is deregulated in many different type of tumors, and thus, Cdks are considered targets for antitumoral therapy. For large screenings of inhibitors the use of purified recombinant Cdks and cyclins is recommended. We report here the current methods to determine their in vitro activity for large screenings of inhibitors.

Key words

Cdk1 Cdk2 Cyclin A Cyclin B Kinase assay 

References

  1. 1.
    Malumbres M, Barbacid M (2005) Mammalian cyclin-dependent kinases. Trends Biochem Sci 30:630–641PubMedCrossRefGoogle Scholar
  2. 2.
    Morgan DO (1997) Cyclin-dependent kinases: engines, clocks, and microprocessors. Annu Rev Cell Dev Biol 13:261–291PubMedCrossRefGoogle Scholar
  3. 3.
    Malumbres M (2014) Cyclin-dependent kinases. Genome Biol 15:122–131PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Rubin SM (2013) Deciphering the retinoblastoma protein phosphorylation code. Trends Biochem Sci 38:12–19PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Lindqvist A, Rodríguez-Bravo V, Medema RH (2009) The decision to enter mitosis: feedback and redundancy in the mitotic entry network. J Cell Biol 185:193–202PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Asghar U, Witkiewicz AK, Turner NC, Knudsen ES (2015) The history and future of targeting cyclin-dependent kinases in cancer therapy. Nat Rev Drug Discov 14:130–146PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    LaBaer J, Garrett MD, Stevenson LF, Slingerland JM, Sandhu C, Chou HS, Fattaey A, Harlow E (1997) New functional activities for the p21 family of CDK inhibitors. Genes Dev 11:847–862PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2016

Authors and Affiliations

  1. 1.Department of Cell Biology, Immunology and NeurosciencesUniversity of BarcelonaBarcelonaSpain
  2. 2.Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS)BarcelonaSpain

Personalised recommendations