Guide to Virtual Screening: Application to the Akt Phosphatase PHLPP

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 819)

Abstract

We present an example-based description of virtual screening (VS) techniques used to identify new regulators of the Akt phosphatase PHLPP (PH domain Leucine repeat Protein Phosphatase). This enzyme opposes the effects of two kinases, Akt and PKC, which play a major role in cell growth and survival. Therefore, PHLPP is a potential therapeutic target in pathophysiologies where these pathways are either repressed, such as in diabetes and cardiovascular diseases, or over-activated as in cancer. To the best of our knowledge, no PHLPP inhibitors have been reported so far in the literature. In this study, we used a combination of chemical and virtual screening techniques that led to the identification of a number of inhibiting compounds with diverse scaffolds. These compounds bind PHLPP and inhibit cell death when tested in cellular assays. We employed GLIDE docking software to screen a library of more than 40,000 compounds selected from the NCI open depository (250,000 compounds) by similarity searches. We compare the efficiency at which we determined binding compounds from the chemical screen, and compare enrichment factors of the virtually discovered compounds over chemical screening.

Key words

Docking Virtual screening PHLPP Akt phosphatase Drug discovery Computer aided drug design 

Notes

Acknowledgments

We thank Professor Alexandra C. Newton for stimulating discussions. This work was supported in part by the Molecular Biophysics Training grant GM08326 (W.S.), the NSF grant MCB-0506593, the NIH grant GM31749, NBCR, CTBP, HHMI, the NSF Supercomputer Centers (J.A.M.), and the Juvenile Diabetes Research Foundation grant 3-2008-478 (E.S.).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Chemistry and Biochemistry, Center for Theoretical Biological PhysicsUniversity of California, San DiegoLa JollaUSA
  2. 2.Department of PharmacologyUniversity of California, San DiegoLa JollaUSA
  3. 3.Department of Chemistry and Biochemistry, Center for Theoretical Biological Physics, Howard Hughes Medical InstituteUniversity of CaliforniaLa JollaUSA
  4. 4.Howard Hughes Medical Institute, Departments of Chemistry and Biochemistry and Pharmacology, Center for Theoretical Biological PhysicsUniversity of California, San DiegoLa JollaUSA

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