Simultaneous in vitro molecular screening of protein-peptide interactions by flow cytometry, using six Bcl-2 family proteins as examples

Nature Protocols

Volumn 6, Issue 7, 2011, Pages 943-952

  Simons, Peter C ^a   Young, Susan M ^a   Carter, Mark B ^a   Waller, Anna ^a   Zhai, Dayong ^b   Reed, John C ^b   Edwards, Bruce S ^a   Sklar, Larry A ^a  

^a UNIVERSITY OF NEW MEXICO SCHOOL OF MEDICINE   (United States)

^b BURNHAM INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

BCL2 RELATED PROTEIN A1; BH3 PROTEIN; CELL PROTEIN; GLUTATHIONE TRANSFERASE; GREEN FLUORESCENT PROTEIN; HYBRID PROTEIN; PROTEIN BCL 2; PROTEIN BCL B; PROTEIN BCL W; PROTEIN BCL XL; PROTEIN MCL 1; UNCLASSIFIED DRUG;

APOPTOSIS; ARTICLE; BINDING ASSAY; CLINICAL PROTOCOL; DIMERIZATION; EXPERIMENTAL DESIGN; FLOW CYTOMETRY; FLUORESCENCE POLARIZATION; GENE OVEREXPRESSION; HYDROPHOBICITY; IN VITRO STUDY; INTERMETHOD COMPARISON; PRIORITY JOURNAL; PROCESS OPTIMIZATION; PROTEIN EXPRESSION; PROTEIN PROTEIN INTERACTION; SCREENING TEST; SENSITIVITY ANALYSIS; SIGNAL TRANSDUCTION; VALIDATION PROCESS;

DIMERIZATION; FLOW CYTOMETRY; FLUORESCENCE; GLUTATHIONE TRANSFERASE; PROTEIN INTERACTION DOMAINS AND MOTIFS; PROTEIN INTERACTION MAPPING; PROTO-ONCOGENE PROTEINS C-BCL-2; RECOMBINANT FUSION PROTEINS;

EID: 79959900394     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2011.339     Document Type: Article

References (18)

1. Gad, S.C. Drug Discovery Handbook (John Wiley and Sons, 2005).
2. Edwards, B.S. et al. Biomolecular screening of formylpeptide receptor ligands with a sensitive, quantitative, high-throughput flow cytometry platform. Nat. Protoc. 1, 59-66 (2006).
3. Tsujimoto, Y., Cossman, J., Jaffe, E. & Croce, C.M. Involvement of the bcl-2 gene in human follicular lymphoma. Science 228, 1440-1443 (1985). (Pubitemid 15041322)
4. Bakhshi, A. et al. Cloning the chromosomal breakpoint of t(14;18) human lymphomas: Clustering around JH on chromosome 14 and near a transcriptional unit on 18. Cell 41, 899-906 (1985). (Pubitemid 15243395)
5. Cleary, M.L., Smith, S.D. & Sklar, J. Cloning and structural analysis of cDNAs for bcl-2 and a hybrid bcl-2/immunoglobulin transcript resulting from the t(14;18) translocation. Cell 47, 19-28 (1986). (Pubitemid 17181529)
6. Youle, R.J. & Strasser, A. The BCL-2 protein family: Opposing activities that mediate cell death. Nat. Rev. Mol. Cell Biol. 9, 47-59 (2008).
7. Sattler, M. et al. Structure of Bcl-xL-Bak peptide complex: Recognition between regulators of apoptosis. Science 275, 983-986 (1997). (Pubitemid 27087715)
8. Chen, L. et al. Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol. Cell 17, 393-403 (2005). (Pubitemid 40193310)
9. Adams, J.M. & Cory, S. The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 26, 1324-1337 (2007). (Pubitemid 46328466)
10. Reed, J.C. & Pellecchia, M. Apoptosis-based therapies for hematologic malignancies. Blood 106, 408-418 (2005). (Pubitemid 40981231)
11. Zhai, D., Jin, C., Satterthwait, A.C. & Reed, J.C. Comparison of chemical inhibitors of antiapoptotic Bcl-2-family proteins. Cell Death. Differ. 13, 1419-1421 (2006). (Pubitemid 44057478)
12. Nakamura, K. et al. Homogeneous time-resolved fluorescence resonance energy transfer assay for measurement of Phox/Bem1p (PB1) domain heterodimerization. J. Biomol. Screen. 13, 396-405 (2008). (Pubitemid 351793988)
13. Owicki, J.C. Fluorescence polarization and anisotropy in high throughput screening: Perspectives and primer. J. Biomol. Screen. 5, 297-306 (2000).
14. Carson, R.T. & Vignali, D.A. Simultaneous quantitation of 15 cytokines using a multiplexed flow cytometric assay. J. Immunol. Methods 227, 41-52 (1999). (Pubitemid 29408703)
15. Curpan, R.F. et al. High-throughput screen for chemical inhibitors of anti-apoptotic Bcl-2 family proteins by multiples flow cytometry. Assay Drug Dev. Technol. (2011).
16. Simons, P.C. et al. Duplexed, bead-based competitive assay for inhibitors of protein kinases. Cytometry A 71, 451-459 (2007). (Pubitemid 47036035)
17. Tessema, M. et al. Glutathione-S-transferase-green fluorescent protein fusion protein reveals slow dissociation from high site density beads and measures free GSH. Cytometry A 69, 326-334 (2006). (Pubitemid 44651345)
18. Bartsch, J.W. et al. An investigation of liquid carryover and sample residual for a high-throughput flow cytometer sample delivery system. Anal. Chem. 76, 3810-3817 (2004). (Pubitemid 38868848)