Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Oncology Letters
      • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Biomedical Reports
      • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • Information for Authors
    • Information for Reviewers
    • Information for Librarians
    • Information for Advertisers
    • Conferences
  • Language Editing
Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • For Authors
    • For Reviewers
    • For Librarians
    • For Advertisers
    • Conferences
  • Language Editing
Login Register Submit
  • This site uses cookies
  • You can change your cookie settings at any time by following the instructions in our Cookie Policy. To find out more, you may read our Privacy Policy.

    I agree
Search articles by DOI, keyword, author or affiliation
Search
Advanced Search
presentation
International Journal of Oncology
Join Editorial Board Propose a Special Issue
Print ISSN: 1019-6439 Online ISSN: 1791-2423
Journal Cover
October-2015 Volume 47 Issue 4

Full Size Image

Cover Legend PDF

Sign up for eToc alerts
Recommend to Library

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

Medicine International

An International Open Access Journal Devoted to General Medicine.

Journal Cover
October-2015 Volume 47 Issue 4

Full Size Image

Cover Legend PDF

Sign up for eToc alerts
Recommend to Library

  • Article
  • Citations
    • Cite This Article
    • Download Citation
    • Create Citation Alert
    • Remove Citation Alert
    • Cited By
  • Similar Articles
    • Related Articles (in Spandidos Publications)
    • Similar Articles (Google Scholar)
    • Similar Articles (PubMed)
  • Download PDF
  • Download XML
  • View XML
Article

A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction

  • Authors:
    • Xiaoqin Fan
    • Jinmei Wei
    • Haiting Xiong
    • Xiaohui Liu
    • Serge Benichou
    • Xuejuan Gao
    • Langxia Liu
  • View Affiliations / Copyright

    Affiliations: Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, Jinan University, Guangzhou 510632, P.R. China, Cochin Institute, INSERM U1016, Centre National de la Recherche Scientifique UMR8104, Université Paris-Descartes, Paris, France
  • Pages: 1485-1493
    |
    Published online on: August 25, 2015
       https://doi.org/10.3892/ijo.2015.3132
  • Expand metrics +
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )
Cited By (CrossRef): 0 citations Loading Articles...

This article is mentioned in:



Abstract

The human immunodeficiency virus (HIV) protein negative factor (Nef) is important for AIDS pathogenesis. An anti-Nef single-domain antibody (sdAb19) derived from camelids has been previously generated and shown to effectively block the physiological functions of Nef in vitro and in vivo in nef-transgenic mice. However, sdAb19 must be ectopically expressed within the target cell to be able to exert its neutralizing effect on Nef, while the extra-cellular administration method turned out to be ineffective. This might suggest a default of the stability or/and deliverability of sdAb19. The identification of small molecule compounds capable of inhibiting the Nef-sdAb19 interaction and mimicking the neutralizing activity of sdAb19 in vivo would therefore be the means of circumventing the problem encountered with sdAb19. Here we describe the development of a high-throughput screening method combining the homogeneous time-resolved fluorescence (HTRF) and the microscale thermophoresis (MST) techniques for the identification of small-molecule compounds inhibiting the Nef-sdAb19 interaction by binding to Nef protein. Eight small-molecule compounds have been selected for their ability to significantly inhibit the Nef-sdAb19 interaction and to bind to Nef. These molecules could be further assessed for their potential of being the Nef-neutralizing agents in the future.
View Figures

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

View References

1 

Greene WC: The brightening future of HIV therapeutics. Nat Immunol. 5:867–871. 2004. View Article : Google Scholar : PubMed/NCBI

2 

Foster JL and Garcia JV: HIV-1 Nef: At the crossroads. Retrovirology. 5:842008. View Article : Google Scholar : PubMed/NCBI

3 

Kestler HW III, Ringler DJ, Mori K, Panicali DL, Sehgal PK, Daniel MD and Desrosiers RC: Importance of the nef gene for maintenance of high virus loads and for development of AIDS. Cell. 65:651–662. 1991. View Article : Google Scholar : PubMed/NCBI

4 

Fackler OT and Baur AS: Live and let die: Nef functions beyond HIV replication. Immunity. 16:493–497. 2002. View Article : Google Scholar : PubMed/NCBI

5 

Shugars DC, Smith MS, Glueck DH, Nantermet PV, Seillier-Moiseiwitsch F and Swanstrom R: Analysis of human immunodeficiency virus type 1 nef gene sequences present in vivo. J Virol. 67:4639–4650. 1993.PubMed/NCBI

6 

Kwak YT, Raney A, Kuo LS, Denial SJ, Temple BR, Garcia JV and Foster JL: Self-association of the Lentivirus protein, Nef. Retrovirology. 7:772010. View Article : Google Scholar : PubMed/NCBI

7 

Cruz NV, Amorim R, Oliveira FE, Speranza FA and Costa LJ: Mutations in the nef and vif genes associated with progression to AIDS in elite controller and slow-progressor patients. J Med Virol. 85:563–574. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Foster JL, Denial SJ, Temple BR and Garcia JV: Mechanisms of HIV-1 Nef function and intracellular signaling. J Neuroimmune Pharmacol. 6:230–246. 2011. View Article : Google Scholar : PubMed/NCBI

9 

Bouchet J, Basmaciogullari SE, Chrobak P, Stolp B, Bouchard N, Fackler OT, Chames P, Jolicoeur P, Benichou S and Baty D: Inhibition of the Nef regulatory protein of HIV-1 by a single-domain antibody. Blood. 117:3559–3568. 2011. View Article : Google Scholar : PubMed/NCBI

10 

Muyldermans S: Single domain camel antibodies: Current status. J Biotechnol. 74:277–302. 2001.PubMed/NCBI

11 

Roovers RC, Laeremans T, Huang L, De Taeye S, Verkleij AJ, Revets H, de Haard HJ and van Bergen en Henegouwen PM: Efficient inhibition of EGFR signaling and of tumour growth by antagonistic anti-EFGR Nanobodies. Cancer Immunol Immunother. 56:303–317. 2007. View Article : Google Scholar

12 

Gueorguieva D, Li S, Walsh N, Mukerji A, Tanha J and Pandey S: Identification of single-domain, Bax-specific intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis. FASEB J. 20:2636–2638. 2006. View Article : Google Scholar : PubMed/NCBI

13 

Hultberg A, Temperton NJ, Rosseels V, Koenders M, Gonzalez-Pajuelo M, Schepens B, Ibañez LI, Vanlandschoot P, Schillemans J, Saunders M, et al: Llama-derived single domain antibodies to build multivalent, superpotent and broadened neutralizing anti-viral molecules. PLoS One. 6:e176652011. View Article : Google Scholar : PubMed/NCBI

14 

Ohmi N, Wingfield JM, Yazawa H and Inagaki O: Development of a homogeneous time-resolved fluorescence assay for high throughput screening to identify Lck inhibitors: Comparison with scintillation proximity assay and streptavidin-coated plate assay. J Biomol Screen. 5:463–470. 2000. View Article : Google Scholar

15 

Degorce F: HTRF®: Pioneering technology for high-throughput screening. Expert Opin Drug Discov. 1:753–764. 2006. View Article : Google Scholar : PubMed/NCBI

16 

Jia Y, Quinn CM, Gagnon AI and Talanian R: Homogeneous time-resolved fluorescence and its applications for kinase assays in drug discovery. Anal Biochem. 356:273–281. 2006. View Article : Google Scholar : PubMed/NCBI

17 

Chen X, Longgood JC, Michnoff C, Wei S, Frantz DE and Bezprozvanny L: High-throughput screen for small molecule inhibitors of Mint1-PDZ domains. Assay Drug Dev Technol. 5:769–783. 2007. View Article : Google Scholar : PubMed/NCBI

18 

Mohamed TM, Zakeri SA, Baudoin F, Wolf M, Oceandy D, Cartwright EJ, Gul S and Neyses L: Optimisation and validation of a high throughput screening compatible assay to identify inhibitors of the plasma membrane calcium ATPase pump - a novel therapeutic target for contraception and malaria. J Pharm Pharm Sci. 16:217–230. 2013.

19 

Lemay J, Maidou-Peindara P, Bader T, Ennifar E, Rain JC, Benarous R and Liu LX: HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in infected cells. Retrovirology. 5:472008. View Article : Google Scholar : PubMed/NCBI

20 

Seidel SA, Dijkman PM, Lea WA, van den Bogaart G, Jerabek-Willemsen M, Lazic A, Joseph JS, Srinivasan P, Baaske P, Simeonov A, et al: Microscale thermophoresis quantifies biomolecular interactions under previously challenging conditions. Methods. 59:301–315. 2013. View Article : Google Scholar :

21 

Fischer WB, Wang YT, Schindler C and Chen CP: Mechanism of function of viral channel proteins and implications for drug development. Int Rev Cell Mol Biol. 294:259–321. 2012. View Article : Google Scholar : PubMed/NCBI

22 

Izzedine H, Launay-Vacher V, Deray G and Hulot JS: Nelfinavir and felodipine: A cytochrome P450 3A4-mediated drug interaction. Clin Pharmacol Ther. 75:362–363. 2004. View Article : Google Scholar : PubMed/NCBI

23 

Idriss HT: Questions regarding the role of protein phosphorylation in HIV replication and anti-acquired immune deficiency syndrome therapy. Saudi Med J. 26:7–10. 2005.PubMed/NCBI

24 

Jakubiec A and Jupin I: Regulation of positive-strand RNA virus replication: The emerging role of phosphorylation. Virus Res. 129:73–79. 2007. View Article : Google Scholar : PubMed/NCBI

25 

Behrend L, Milne DM, Stöter M, Deppert W, Campbell LE, Meek DW and Knippschild U: IC261, a specific inhibitor of the protein kinases casein kinase 1-delta and -epsilon, triggers the mitotic checkpoint and induces p53-dependent postmitotic effects. Oncogene. 19:5303–5313. 2000. View Article : Google Scholar : PubMed/NCBI

26 

Yang WS and Stockwell BR: Inhibition of casein kinase 1-epsilon induces cancer-cell-selective, PERIOD2-dependent growth arrest. Genome Biol. 9:R922008. View Article : Google Scholar : PubMed/NCBI

27 

Zhou YX, Karle S, Taguchi H, Planque S, Nishiyama Y and Paul S: Prospects for immunotherapeutic proteolytic antibodies. J Immunol Methods. 269:257–268. 2002. View Article : Google Scholar : PubMed/NCBI

28 

Evans TR and Kaye SB: Retinoids: Present role and future potential. Br J Cancer. 80:1–8. 1999. View Article : Google Scholar : PubMed/NCBI

29 

Kaleagasioglu F, Doepner G, Biesalski HK and Berger MR: Antiproliferative activity of retinoic acid and some novel retinoid derivatives in breast and colorectal cancer cell lines of human origin. Arzneimittelforschung. 43:487–490. 1993.PubMed/NCBI

30 

Hua S, Kittler R and White KP: Genomic antagonism between retinoic acid and estrogen signaling in breast cancer. Cell. 137:1259–1271. 2009. View Article : Google Scholar : PubMed/NCBI

31 

He JC, Lu TC, Fleet M, Sunamoto M, Husain M, Fang W, Neves S, Chen Y, Shankland S, Iyengar R, et al: Retinoic acid inhibits HIV-1-induced podocyte proliferation through the cAMP pathway. J Am Soc Nephrol. 18:93–102. 2007. View Article : Google Scholar

32 

Lu TC, Wang Z, Feng X, Chuang P, Fang W, Chen Y, Neves S, Maayan A, Xiong H, Liu Y, et al: Retinoic acid utilizes CREB and USF1 in a transcriptional feed-forward loop in order to stimulate MKP1 expression in human immunodeficiency virus-infected podocytes. Mol Cell Biol. 28:5785–5794. 2008. View Article : Google Scholar : PubMed/NCBI

Related Articles

  • Abstract
  • View
  • Download
  • Twitter
Copy and paste a formatted citation
Spandidos Publications style
Fan X, Wei J, Xiong H, Liu X, Benichou S, Gao X and Liu L: A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction. Int J Oncol 47: 1485-1493, 2015.
APA
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., & Liu, L. (2015). A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction. International Journal of Oncology, 47, 1485-1493. https://doi.org/10.3892/ijo.2015.3132
MLA
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., Liu, L."A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction". International Journal of Oncology 47.4 (2015): 1485-1493.
Chicago
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., Liu, L."A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction". International Journal of Oncology 47, no. 4 (2015): 1485-1493. https://doi.org/10.3892/ijo.2015.3132
Copy and paste a formatted citation
x
Spandidos Publications style
Fan X, Wei J, Xiong H, Liu X, Benichou S, Gao X and Liu L: A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction. Int J Oncol 47: 1485-1493, 2015.
APA
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., & Liu, L. (2015). A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction. International Journal of Oncology, 47, 1485-1493. https://doi.org/10.3892/ijo.2015.3132
MLA
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., Liu, L."A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction". International Journal of Oncology 47.4 (2015): 1485-1493.
Chicago
Fan, X., Wei, J., Xiong, H., Liu, X., Benichou, S., Gao, X., Liu, L."A homogeneous time-resolved fluorescence-based high-throughput screening for discovery of inhibitors of Nef-sdAb19 interaction". International Journal of Oncology 47, no. 4 (2015): 1485-1493. https://doi.org/10.3892/ijo.2015.3132
Follow us
  • Twitter
  • LinkedIn
  • Facebook
About
  • Spandidos Publications
  • Careers
  • Cookie Policy
  • Privacy Policy
How can we help?
  • Help
  • Live Chat
  • Contact
  • Email to our Support Team