The PI3Ks (phosphatidylinositol 3-kinases) regulate cellular signalling networks that are involved in processes linked to the survival, growth, proliferation, metabolism and specialized differentiated functions of cells. The subversion of this network is common in cancer and has also been linked to disorders of inflammation. The elucidation of the physiological function of PI3K has come from pharmacological studies, which use the enzyme inhibitors Wortmannin and LY294002, and from PI3K genetic knockout models of the effects of loss of PI3K function. Several reports have shown that LY294002 is not exclusively selective for the PI3Ks, and could in fact act on other lipid kinases and additional apparently unrelated proteins. Since this inhibitor still remains a drug of choice in numerous PI3K studies (over 500 in the last year), it is important to establish the precise specificity of this compound. We report here the use of a chemical proteomic strategy in which an analogue of LY294002, PI828, was immobilized onto epoxy-activated Sepharose beads. This affinity material was then used as a bait to fish-out potential protein targets from cellular extracts. Proteins with high affinity for immobilized PI828 were separated by one-dimensional gel electrophoresis and identified by liquid chromatography–tandem MS. The present study reveals that LY294002 not only binds to class I PI3Ks and other PI3K-related kinases, but also to novel targets seemingly unrelated to the PI3K family.
Skip Nav Destination
Article navigation
May 2007
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
April 26 2007
Exploring the specificity of the PI3K family inhibitor LY294002
Severine I. Gharbi;
Severine I. Gharbi
*Ludwig Institute for Cancer Research, Proteomics Unit, Cruciform Building, Gower Street, London WCE1 6BT, U.K.
Search for other works by this author on:
Marketa J. Zvelebil;
Marketa J. Zvelebil
†Ludwig Institute for Cancer Research, Bioinformatics Group, 91 Riding House Street, London W1W 7BS, U.K.
Search for other works by this author on:
Stephen J. Shuttleworth;
Stephen J. Shuttleworth
‡Plramed, 957 Buckingham Avenue, Slough, Berkshire SL1 4NL, U.K.
Search for other works by this author on:
Tim Hancox;
Tim Hancox
‡Plramed, 957 Buckingham Avenue, Slough, Berkshire SL1 4NL, U.K.
Search for other works by this author on:
Nahid Saghir;
Nahid Saghir
‡Plramed, 957 Buckingham Avenue, Slough, Berkshire SL1 4NL, U.K.
Search for other works by this author on:
John F. Timms;
John F. Timms
*Ludwig Institute for Cancer Research, Proteomics Unit, Cruciform Building, Gower Street, London WCE1 6BT, U.K.
§Transitional Research Laboratory, Institute of Women's Health, University College London, Huntley Street, London WC1E 6DH, U.K.
Search for other works by this author on:
Michael D. Waterfield
Michael D. Waterfield
1
*Ludwig Institute for Cancer Research, Proteomics Unit, Cruciform Building, Gower Street, London WCE1 6BT, U.K.
‡Plramed, 957 Buckingham Avenue, Slough, Berkshire SL1 4NL, U.K.
1To whom correspondence should be addressed (email mikew@ludwig.ucl.ac.uk).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
September 29 2006
Revision Received:
February 12 2007
Accepted:
February 15 2007
Accepted Manuscript online:
February 15 2007
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2007 Biochemical Society
2007
Biochem J (2007) 404 (1): 15–21.
Article history
Received:
September 29 2006
Revision Received:
February 12 2007
Accepted:
February 15 2007
Accepted Manuscript online:
February 15 2007
Citation
Severine I. Gharbi, Marketa J. Zvelebil, Stephen J. Shuttleworth, Tim Hancox, Nahid Saghir, John F. Timms, Michael D. Waterfield; Exploring the specificity of the PI3K family inhibitor LY294002. Biochem J 15 May 2007; 404 (1): 15–21. doi: https://doi.org/10.1042/BJ20061489
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.