Phosphoproteomic profiling of tumor tissues identifies HSP27 Ser82 phosphorylation as a robust marker of early ischemia

Scientific Reports
Muhammad Saddiq ZahariAkhilesh Pandey

Abstract

Delays between tissue collection and tissue fixation result in ischemia and ischemia-associated changes in protein phosphorylation levels, which can misguide the examination of signaling pathway status. To identify a biomarker that serves as a reliable indicator of ischemic changes that tumor tissues undergo, we subjected harvested xenograft tumors to room temperature for 0, 2, 10 and 30 minutes before freezing in liquid nitrogen. Multiplex TMT-labeling was conducted to achieve precise quantitation, followed by TiO2 phosphopeptide enrichment and high resolution mass spectrometry profiling. LC-MS/MS analyses revealed phosphorylation level changes of a number of phosphosites in the ischemic samples. The phosphorylation of one of these sites, S82 of the heat shock protein 27 kDa (HSP27), was especially abundant and consistently upregulated in tissues with delays in freezing as short as 2 minutes. In order to eliminate effects of ischemia, we employed a novel cryogenic biopsy device which begins freezing tissues in situ before they are excised. Using this device, we showed that the upregulation of phosphorylation of S82 on HSP27 was abrogated. We thus demonstrate that our cryogenic biopsy device can eliminate ischemia-induced phosp...Continue Reading

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Citations

Jul 3, 2019·Journal of Bacteriology·Landon J GetzNikhil A Thomas
Oct 23, 2019·Signal Transduction and Targeted Therapy·Hang SongQiuhui Pan
Apr 26, 2020·Gastric Cancer : Official Journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association·Niraj BabuAditi Chatterjee
Oct 11, 2020·Signal Transduction and Targeted Therapy·Hang SongQiuhui Pan

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Methods Mentioned

BETA
xenograft
biopsy
xenografts
electrophoresis

Software Mentioned

SEQUEST
Mascot
Proteome Discoverer

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