Selective Labeling and Identification of the Tumor Cell Proteome of Pancreatic Cancer In Vivo

• DOI: 10.1021/acs.jproteome.0c00666
• Publication Date: 2020-12-09T18:04:28Z
• Authors: Nancy G. Azizian; Delaney K. Sullivan; Litong Nie; Sammy Pardo; Dana Molleur; Junjie Chen; Susan T. Weintraub; Yulin Li
• Keywords: Proteome; Bioorthogonal Chemistry; Tumor initiation; Tumor progression

Pancreatic ductal adenocarcinoma (PDAC) is among the deadliest cancers. Dissecting tumor cell proteome from that of non-tumor cells in PDAC bulk critical for tumorigenesis studies, biomarker discovery, and development therapeutics. However, investigating has proven evasive due to tumor's extremely complex cellular composition. To circumvent this technical barrier, we have combined bioorthogonal noncanonical amino acid tagging (BONCAT) data-independent acquisition mass spectrometry (DIA-MS) an orthotopic model specifically identify vivo. Utilizing cell-specific expression a mutant tRNA synthetase transgene, approach provides with exclusive ability incorporate azide-bearing methionine analogue into newly synthesized proteins. The azide-tagged subsequently enriched purified via reaction then identified quantified using DIA-MS. Applying workflow model, thousands proteins expressed by cells. Furthermore, comparing proteomes, showed can distinctly differentiate produced those within microenvironment. Our study, first time, reveals under physiological conditions, providing broad applications tumorigenesis, therapeutics, studies various human