Literature DB >> 34679246

Protocols for Characterization of Cdk5 Kinase Activity.

Anita Terse1, Niranjana Amin2, Bradford Hall1, Manju Bhaskar2, Binukumar B K3, Elias Utreras4, Tej K Pareek5, Harish Pant2, Ashok B Kulkarni1.   

Abstract

Cyclin-dependent kinases (Cdks) are generally known to be involved in controlling the cell cycle, but Cdk5 is a unique member of this protein family for being most active in post-mitotic neurons. Cdk5 is developmentally important in regulating neuronal migration, neurite outgrowth, and axon guidance. Cdk5 is enriched in synaptic membranes and is known to modulate synaptic activity. Postnatally, Cdk5 can also affect neuronal processes such as dopaminergic signaling and pain sensitivity. Dysregulated Cdk5, in contrast, has been linked to neurodegenerative disorders such as Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS). Despite primarily being implicated in neuronal development and activity, Cdk5 has lately been linked to non-neuronal functions including cancer cell growth, immune responses, and diabetes. Since Cdk5 activity is tightly regulated, a method for measuring its kinase activity is needed to fully understand the precise role of Cdk5 in developmental and disease processes. This article includes methods for detecting Cdk5 kinase activity in cultured cells or tissues, identifying new substrates, and screening for new kinase inhibitors. Furthermore, since Cdk5 shares homology and substrate specificity with Cdk1 and Cdk2, the Cdk5 kinase assay can be used, with modification, to measure the activity of other Cdks as well.
© 2021 Wiley Periodicals LLC. This article has been contributed to by US Government employees and their work is in the public domain in the USA. Basic Protocol 1: Measuring Cdk5 activity from protein lysates Support Protocol 1: Immunoprecipitation of Cdk5 using Dynabeads Alternate Protocol: Non-radioactive protocols to measure Cdk5 kinase activity Support Protocol 2: Western blot analysis for the detection of Cdk5, p35, and p39 Support Protocol 3: Immunodetection analysis for Cdk5, p35, and p39 Support Protocol 4: Genetically engineered mice (+ and - controls) Basic Protocol 2: Identifying new Cdk5 substrates and kinase inhibitors. © 2021 Wiley Periodicals LLC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Entities:  

Keywords:  Cdk5; kinase assay; p25; p35; p39

Mesh:

Substances:

Year:  2021        PMID: 34679246      PMCID: PMC8555461          DOI: 10.1002/cpz1.276

Source DB:  PubMed          Journal:  Curr Protoc        ISSN: 2691-1299


  92 in total

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Authors:  Zhenxin Wang; Raphaël Lévy; David G Fernig; Mathias Brust
Journal:  J Am Chem Soc       Date:  2006-02-22       Impact factor: 15.419

2.  Identification of the 23 kDa subunit of tau protein kinase II as a putative activator of cdk5 in bovine brain.

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Journal:  FEBS Lett       Date:  1994-04-04       Impact factor: 4.124

3.  Calpain-mediated cleavage of the cyclin-dependent kinase-5 activator p39 to p29.

Authors:  Holger Patzke; Li-Huei Tsai
Journal:  J Biol Chem       Date:  2002-01-09       Impact factor: 5.157

4.  Structure and regulation of the CDK5-p25(nck5a) complex.

Authors:  C Tarricone; R Dhavan; J Peng; L B Areces; L H Tsai; A Musacchio
Journal:  Mol Cell       Date:  2001-09       Impact factor: 17.970

5.  Enzyme colorimetric assay using unmodified silver nanoparticles.

Authors:  Hui Wei; Chaogui Chen; Bingyan Han; Erkang Wang
Journal:  Anal Chem       Date:  2008-07-29       Impact factor: 6.986

6.  Generation of transgenic mice.

Authors:  Andrew Cho; Naoto Haruyama; Ashok B Kulkarni
Journal:  Curr Protoc Cell Biol       Date:  2009-03

7.  Phosphorylation of cyclin-dependent kinase 5 (Cdk5) at Tyr-15 is inhibited by Cdk5 activators and does not contribute to the activation of Cdk5.

Authors:  Hiroyuki Kobayashi; Taro Saito; Ko Sato; Kotaro Furusawa; Tomohisa Hosokawa; Koji Tsutsumi; Akiko Asada; Shinji Kamada; Toshio Ohshima; Shin-ichi Hisanaga
Journal:  J Biol Chem       Date:  2014-05-28       Impact factor: 5.157

8.  Phosphorylation of the Transient Receptor Potential Ankyrin 1 by Cyclin-dependent Kinase 5 affects Chemo-nociception.

Authors:  Bradford E Hall; Michaela Prochazkova; Matthew R Sapio; Paul Minetos; Natalya Kurochkina; B K Binukumar; Niranjana D Amin; Anita Terse; John Joseph; Stephen J Raithel; Andrew J Mannes; Harish C Pant; Man-Kyo Chung; Michael J Iadarola; Ashok B Kulkarni
Journal:  Sci Rep       Date:  2018-01-19       Impact factor: 4.379

9.  MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI).

Authors:  Itxaso Bellón-Echeverría; Jean-Philippe Carralot; Andrea Araujo Del Rosario; Stephanie Kueng; Harald Mauser; Georg Schmid; Ralf Thoma; Imre Berger
Journal:  Sci Rep       Date:  2018-03-23       Impact factor: 4.379

10.  Activity and expression pattern of cyclin-dependent kinase 5 in the embryonic mouse nervous system.

Authors:  L H Tsai; T Takahashi; V S Caviness; E Harlow
Journal:  Development       Date:  1993-12       Impact factor: 6.868

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