Talk Title: Pulse Azidohomoalanine (AHA) Labeling in Mammals (PALM) analysis for global analysis of newly-synthesized proteins in animal models of disease
John R. Yates is the Ernest W. Hahn Professor in the Department of Molecular Medicine and Neurobiology at The Scripps Research Institute. His research interests include development of integrated methods for tandem mass spectrometry analysis of protein mixtures, bioinformatics using mass spectrometry data, and biological studies involving proteomics. He is the lead inventor of the SEQUEST software for correlating tandem mass spectrometry data to sequences in the database and developer of the shotgun proteomics technique for the analysis of protein mixtures. He has received the American Society for Mass Spectrometry research award, the Pehr Edman Award in Protein Chemistry, the American Society for Mass Spectrometry Biemann Medal, the HUPO Distinguished Achievement Award in Proteomics, Herbert Sober Award from the ASBMB, and the Christian Anfinsen Award from The Protein Society, the 2015 ACS’s Analytical Chemistry award and 2015 The Ralph N. Adams Award in Bioanalytical Chemistry. Dr. Yates is the Editor in Chief at the Journal of Proteome Research.
Pulse Azidohomoalanine (AHA) Labeling in Mammals (PALM) analysis for global analysis of newly-synthesized proteins in animal models of disease
John R. Yates, III, Yuanhui Ma, Daniel McClatchy
Identifying changes in animal models of disease at the earliest time point prior to pathology is desired because these alterations are more likely to cause the pathology. At these early time points alterations in protein expression may be difficult to identify hidden by the overwhelming static proteome. One possible solution is to identify only newly synthesized proteins (NSP) within a discrete time period. We developed PALM analysis to identify and quantify NSP from rodent tissues by mass spectrometry. When introduced by Tirrell and Schumann it was also referred to as BONCAT. A rodent diet was developed where methionine was replaced with AHA. Mice were given the PALM diet for different time periods and multiple tissues were tested for incorporation of AHA. To quantify the NSP, a heavy biotin-alkyne and heavy AHA were synthesized. The use of these reagents will be discussed in animal models and cell culture experiments.