A 96-well enzyme-linked immunosorbent assay was developed to discover compounds that inhibit the binding of the Leu-X-Cys-X-Glu (LXCXE) motif to the retinoblastoma tumor suppressor protein (pRB). The assay uses a LXCXE-containing multiple antigenic peptide (MAP) which is immobilized on a microtiter plate. A truncated form of pRB is added and the amount bound detected by a monoclonal antibody. This rapid assay was employed in high throughput screening of crude natural product extracts and discrete compounds.
References
1.
Weinberg, R.A. (1996). How cancer arises. Sci. Am.275:62.
2.
Oliff, A., Gibbs, J.B., McCormick, F. (1996). New molecular targets for cancer therapy. Sci. Am.275:144.
3.
Weinberg, R.A. (1995). The retinoblastoma protein and cell cycle control. Cell81:323.
4.
Paggi, M.G., Baldi, A., Bonetto, F., Giordano, A. (1996). Retinoblastoma protein family in cell cycle and cancer: A review. J. Cell. Biochem.62:418.
5.
Sanchez, I., Dynlacht, B.D. (1996). Transcriptional control of the cell cycle. Curr. Op. Cell. Biol.8:318.
6.
Jansen-Dirr, P. (1996). How viral oncogenes make the cell cycle. Trends Genet.12:270.
7.
Taya, Y. (1997). RB kinases and RB-binding proteins: New points of view. Trends Biochem Sci.22:14.
8.
Pfister, H. (1996). The role of human papillomavirus in anogenital cancer. Obstet. Gynecol. Clin. North Am.23:579.
9.
Hall, M., Peters, G. (1996). Genetic alterations of cyclins, cyclindependent kinases, and Cdk inhibitors in human cancer. Adv. Cancer Res.68:67.
10.
Smith, D.B., Corcoran, L.M. (1991). Expression and purification of glutathione-S-transferase fusion proteins. In Current Protocols in Molecular Biology. F.A. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, and K. Struhl (eds.), pp. 16.7.1-16.7.8. Greene Publishing and Wiley-Interscience, New York.
11.
Bradford, M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem.72:248.
12.
Jones, R.E., Wegrzyn, R.J., Patrick, D.R., Balishin, N.L., Vuocolo, G.A., Riemen, M.W., Defeo-Jones, D., Garsky, V.M., Heimbrook, D.C., Oliff, A. (1990). Identification of HPV-16 E7 peptides that are potent antagonists of E7 binding to the retinoblastoma suppressor protein. J. Biol. Chem.265:12782.
13.
Jones, R.E., Heimbrook, D.C., Huber, H.E., Wegrzyn, R.J., Rotberg, N.S., Stauffer, K.J., Lumma, P.K., Oliff, A. (1992). Specific N-methylations of HPV-16 peptides alter binding to the retinoblastoma suppressor protein. J. Biol. Chem.267:908.
14.
Huber, H.E., Koblan, K.S., Heimbrook, D.C. (1994). Protein-protein interactions as therapeutic targets for cancer. Curr. Med. Chem.1:13.
15.
Patrick, D., Oliff, A., Heimbrook, D.C. (1994). Identification of a novel retinoblastoma gene product binding site on human papillomavirus type 16 E7 protein. J. Biol. Chem.269:6842.
16.
Tam, J.P. (1988). Synthetic peptide vaccine design: Synthesis and properties of a high-density multiple antigenic peptide system. Proc. Natl. Acad. Sci. USA85:5409.
17.
Tam, J.P. (1996). Recent advances in multiple antigen peptides. J. Immunol. Meth.196:17.
18.
Palmieri, G., Cassani, G., Fassina, G. (1995). Peptide immobilization on calcium alginate beads-applications to antibody purification and assay. J. Chromatog. B664:127.
19.
Hayakawa, Y., Sohda, K.-Y., Shin-ya, K., Hidaka, T., Seto, H. (1995). Anguinomycins C and D, new antitumor antibiotics with selective cytotoxicity against transformed cells. J. Antibiot.48:954.
20.
Hayakawa, Y., Sohda, K.-Y., Seto, H. (1995). Selective cytotoxicity of mycophenolic acid against transformed cells. J. Antibiot.48:1182.
