Bioorthogonal Labeling of Human Prostate Cancer Tissue Slice Cultures for Glycoproteomics.

Autor: Spiciarich DR; College of Chemistry, University of California, Berkeley, Berkeley, CA, 94720, USA., Nolley R; Department of Urology, Stanford University School of Medicine, Stanford, CA, 94305, USA., Maund SL; Department of Urology, Stanford University School of Medicine, Stanford, CA, 94305, USA., Purcell SC; College of Chemistry, University of California, Berkeley, Berkeley, CA, 94720, USA., Herschel J; Department of Mathematics, California State University, East Bay Hayward, CA, 94542, USA., Iavarone AT; QB3/Chemistry Mass Spectrometry Facility, UC Berkeley, Berkeley, CA, 94720, USA., Peehl DM; Department of Urology, Stanford University School of Medicine, Stanford, CA, 94305, USA., Bertozzi CR; Department of Chemistry, Stanford University, Stanford, CA, 94305-4401, USA.; Howard Hughes Medical Institute, USA.
Jazyk: angličtina
Zdroj: Angewandte Chemie (International ed. in English) [Angew Chem Int Ed Engl] 2017 Jul 24; Vol. 56 (31), pp. 8992-8997. Date of Electronic Publication: 2017 Jun 26.
DOI: 10.1002/anie.201701424
Abstrakt: Sialylated glycans are found at elevated levels in many types of cancer and have been implicated in disease progression. However, the specific glycoproteins that contribute to the cancer cell-surface sialylation are not well characterized, specifically in bona fide human disease tissue. Metabolic and bioorthogonal labeling methods have previously enabled the enrichment and identification of sialoglycoproteins from cultured cells and model organisms. Herein, we report the first application of this glycoproteomic platform to human tissues cultured ex vivo. Both normal and cancerous prostate tissues were sliced and cultured in the presence of the azide-functionalized sialic acid biosynthetic precursor Ac 4 ManNAz. The compound was metabolized to the azidosialic acid and incorporated into cell surface and secreted sialoglycoproteins. Chemical biotinylation followed by enrichment and mass spectrometry led to the identification of glycoproteins that were found at elevated levels or uniquely in cancerous prostate tissue. This work therefore extends the use of bioorthogonal labeling strategies to problems of clinical relevance.
(© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
Databáze: MEDLINE
Externí odkaz: