PROJECT SUMMARY/ABSTRACT The Proteomics Resource Laboratory Shared Resource (PRL) was established 10 years ago under the direction of Beatrix Ueberheide, PhD, Associate Professor of Biochemistry and Molecular Pharmacology and Neurology. She has over 20 years of experience in mass spectrometry (MS) with over 100 publications and is a well-recognized expert in biological MS. The PRL mission is to aid scientific discovery by Perlmutter Cancer Center (PCC) members by providing access to specialized expertise and state-of-the-art instrumentation and techniques for MS analysis of proteins. PRL provides advice and full-service MS-based proteomics analysis in a highly collaborative manner to foster cutting-edge research by PCC members. PRL services are critical to the four PCC Research Programs as proteomic analysis is central to much of basic, translational, and clinical cancer research. During the previous funding period, 57 PCC members used PRL, 25 PCC publications included PRL data, and 24 of these had PRL co-authors. PRL consults on experimental design and provides full support of downstream data analysis, often using innovative data analysis pipelines. Significant economies of scale are achieved by this centralization of resources. To provide for future needs of PCC members and remain state-of- the-art, PRL is continuously improving, developing, and implementing new services via its highly trained staff and updating instrumentation via generous institutional support. PRL is overseen by the Division of Advanced Research Services, solicits frequent surveys from PCC members, and consults an Advisory Board. PRL achieves its mission via four Specific Aims: 1) To provide MS-based state-of-the-art proteomics technologies, complete downstream data analysis and storage support to PCC members. These technologies include, but are not limited to, quantitative mass spectrometry using label and label-free strategies, identification and quantification of post-translational-modifications, and characterization of protein complexes; 2) To provide highly customized project support tailored to the specific research questions and needs of individual PCC members, including method(s) development and/or unique data analysis strategies for these applications, 3) To continuously implement and develop novel proteomics techniques and, as needed, secure new instrumentation to ensure continued availability of state-of-the-art proteomics technology to PCC members, 4) To provide education to graduate students, post-doctoral researchers, and principal investigators on proteomics techniques by participating in graduate student teaching, departmental seminars, and extensive pre- and post-project consultation and collaboration.