A Rapid Library Screen for Tailoring -Peptide Structure and Function
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- 作者：Joshua A. Kritzer ; Nathan W. Luedtke ; Elizabeth A. Harker ; and Alanna Schepartz
- 刊名：Journal of the American Chemical Society
- 出版年：2005
- 出版时间：October 26, 2005
- 年：2005
- 卷：127
- 期：42
- 页码：14584 - 14585
- 全文大小：102K
- 年卷期：v.127,no.42(October 26, 2005)
- ISSN：1520-5126
文摘
Recently we described a 
-decapeptide (53-1) that folds into a 14-helix in aqueous solution, binds the oncoprotein hDM2 with submicromolar affinity, and inhibits the interaction of hDM2 with a peptide derived from the activation domain of p53 (p53AD). The solution structure of 53-1 in CD3OH revealed an unexpected C-terminal unwinding that staggers the side chains comprising the hDM2 recognition epitope to better mimic those of p53AD. The structure-function relationship implied by this distortion suggested that a library of 53-1 analogues possessing diversity along a nonrecognition face might contain molecules possessing greater affinity for hDM2. Here we describe (1) -peptide synthesis protocols that produce high quality one-bead-one--peptide libraries suitable for on-bead screening without purification, (2) a versatile, scalable on-bead screen, and (3) a simple tandem mass spectrometry (MS/MS) decoding method. Using this procedure, we identified 53-1 analogues with improved structural and functional properties.
-decapeptide (53-1) that folds into a 14-helix in aqueous solution, binds the oncoprotein hDM2 with submicromolar affinity, and inhibits the interaction of hDM2 with a peptide derived from the activation domain of p53 (p53AD). The solution structure of 53-1 in CD3OH revealed an unexpected C-terminal unwinding that staggers the side chains comprising the hDM2 recognition epitope to better mimic those of p53AD. The structure-function relationship implied by this distortion suggested that a library of 53-1 analogues possessing diversity along a nonrecognition face might contain molecules possessing greater affinity for hDM2. Here we describe (1) -peptide synthesis protocols that produce high quality one-bead-one--peptide libraries suitable for on-bead screening without purification, (2) a versatile, scalable on-bead screen, and (3) a simple tandem mass spectrometry (MS/MS) decoding method. Using this procedure, we identified 53-1 analogues with improved structural and functional properties.