Novel Bcl-2 Homology-3 Domain-like Sequences Identified from Screening Randomized Peptide Libraries for Inhibitors of the Pro-survival Bcl-2 Proteins Export

@article{citeulike:6044036, abstract = {10.1074/jbc.M109.048009 Interactions between Bcl-2 homology-3 (BH3)-only proteins and their pro-survival Bcl-2 family binding partners initiate the intrinsic apoptosis pathway. These interactions are mediated by a short helical motif, the BH3 domain, on the BH3-only protein, which inserts into a hydrophobic groove on the pro-survival molecule. To identify novel peptidic ligands that bind Mcl-1, a pro-survival protein relative of Bcl-2, both human and mouse Mcl-1 were screened against large randomized phage-displayed peptide libraries. We identified a number of 16-mer peptides with sub-micromolar affinity that were highly selective for Mcl-1, as well as being somewhat selective for the species of Mcl-1 (human or mouse) against which the library was panned. Interestingly, these sequences all strongly resembled natural BH3 domain sequences. By switching residues within the best of the human Mcl-1-binding sequences, or extending beyond the core sequence identified, we were able to alter the pro-survival protein interaction profile of this peptide such that it now bound all members tightly and was a potent killer when introduced into cells. Introduction of an amide lock constraint within this sequence also increased its helicity and binding to pro-survival proteins. These data provide new insights into the determinants of BH3 domain:pro-survival protein affinity and selectivity.}, author = {Lee, Erinna F. and Fedorova, Anna and Zobel, Kerry and Boyle, Michelle J. and Yang, Hong and Perugini, Matthew A. and Colman, Peter M. and Huang, David C. S. and Deshayes, Kurt and Fairlie, W. Douglas}, citeulike-article-id = {6044036}, citeulike-linkout-0 = {http://dx.doi.org/10.1074/jbc.M109.048009}, citeulike-linkout-1 = {http://www.jbc.org/content/284/45/31315.abstract}, citeulike-linkout-2 = {http://www.jbc.org/content/284/45/31315.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/19748896}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=19748896}, day = {6}, doi = {10.1074/jbc.M109.048009}, journal = {Journal of Biological Chemistry}, keywords = {bcl-2, bh3, lee09pdf, libraries, peptide}, month = {November}, number = {45}, pages = {31315--31326}, posted-at = {2009-10-30 19:22:23}, priority = {2}, title = {Novel Bcl-2 Homology-3 Domain-like Sequences Identified from Screening Randomized Peptide Libraries for Inhibitors of the Pro-survival Bcl-2 Proteins}, url = {http://dx.doi.org/10.1074/jbc.M109.048009}, volume = {284}, year = {2009} }

TY - JOUR ID - citeulike:6044036 L3 - citeulike-article-id:6044036 N2 - 10.1074/jbc.M109.048009 Interactions between Bcl-2 homology-3 (BH3)-only proteins and their pro-survival Bcl-2 family binding partners initiate the intrinsic apoptosis pathway. These interactions are mediated by a short helical motif, the BH3 domain, on the BH3-only protein, which inserts into a hydrophobic groove on the pro-survival molecule. To identify novel peptidic ligands that bind Mcl-1, a pro-survival protein relative of Bcl-2, both human and mouse Mcl-1 were screened against large randomized phage-displayed peptide libraries. We identified a number of 16-mer peptides with sub-micromolar affinity that were highly selective for Mcl-1, as well as being somewhat selective for the species of Mcl-1 (human or mouse) against which the library was panned. Interestingly, these sequences all strongly resembled natural BH3 domain sequences. By switching residues within the best of the human Mcl-1-binding sequences, or extending beyond the core sequence identified, we were able to alter the pro-survival protein interaction profile of this peptide such that it now bound all members tightly and was a potent killer when introduced into cells. Introduction of an amide lock constraint within this sequence also increased its helicity and binding to pro-survival proteins. These data provide new insights into the determinants of BH3 domain:pro-survival protein affinity and selectivity. IS - 45 JF - Journal of Biological Chemistry EP - 31326 TI - Novel Bcl-2 Homology-3 Domain-like Sequences Identified from Screening Randomized Peptide Libraries for Inhibitors of the Pro-survival Bcl-2 Proteins VL - 284 SP - 31315 KW - bcl-2 KW - bh3 KW - lee09pdf KW - libraries KW - peptide AU - Lee, Erinna AU - Fedorova, Anna AU - Zobel, Kerry AU - Boyle, Michelle AU - Yang, Hong AU - Perugini, Matthew AU - Colman, Peter AU - Huang, David AU - Deshayes, Kurt AU - Fairlie, Douglas PY - 2009/11/06/ UR - http://dx.doi.org/10.1074/jbc.M109.048009 ER -