non-natural and photo-reactive amino acids as biochemical probes of immune function非天然的和photo-reactive氨基酸生物化学探针的免疫功能.pdfVIP

Non-Natural and Photo-Reactive Amino Acids as Biochemical Probes of Immune Function ´ ˜ 1. 1,2. 1 1,2 1,2 Marta Gomez-Nunez , Kurtis J. Haro , Tao Dao , Deming Chau , Annie Won , Sindy Escobar- 1,2 1 1 1 1 Alvarez , Victoriya Zakhaleva , Tatyana Korontsvit , David Y. Gin , David A. Scheinberg * 1 Molecular Pharmacology and Chemistry Program, Memorial Sloan-Kettering Cancer Center, New York, New York, United States of America, 2 Weill Cornell Graduate School of Medical Sciences, New York, New York, United States of America Abstract Wilms tumor protein (WT1) is a transcription factor selectively overexpressed in leukemias and cancers; clinical trials are underway that use altered WT1 peptide sequences as vaccines. Here we report a strategy to study peptide-MHC interactions by incorporating non-natural and photo-reactive amino acids into the sequence of WT1 peptides. Thirteen WT1 peptides sequences were synthesized with chemically modified amino acids (via fluorination and photo-reactive group additions) at MHC and T cell receptor binding positions. Certain new non-natural peptide analogs could stabilize MHC class I molecules better than the native sequences and were also able to elicit specific T-cell responses and sometimes cytotoxicity to leukemia cells. Two photo-reactive peptides, also modified with a biotin handle for pull-down studies, formed covalent interactions with MHC molecules on live cells and provided kinetic data showing the rapid clearance of the peptide-MHC complex. Despite ‘‘infinite affinity’’ provided by the covalent pepti