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Am J Physiol Renal Physiol. 2019 Jun 1;316(6):F1236-F1243. doi: 10.1152/ajprenal.00387.2018. Epub 2019 Apr 17.

Quantitative proteomic analysis of a genetically induced prostate inflammation mouse model via custom 4-plex DiLeu isobaric labeling.

Author information

1
School of Pharmacy, University of Wisconsin-Madison , Madison, Wisconsin.
2
Molecular and Environmental Toxicology Center, University of Wisconsin-Madison , Madison, Wisconsin.
3
Department of Chemistry, University of Wisconsin-Madison , Madison, Wisconsin.
4
School of Veterinary Medicine, University of Wisconsin-Madison , Madison, Wisconsin.
5
George M. O'Brien Center of Research Excellence, University of Wisconsin-Madison , Madison, Wisconsin.
6
Department of Biological Sciences, University of Maryland-Baltimore County , Baltimore, Maryland.
7
Department of Pathology, Johns Hopkins University School of Medicine , Baltimore, Maryland.
8
University of Maryland Marlene and Stewart Greenebaum Cancer Center , Baltimore, Maryland.
9
Department of Urology, University of Wisconsin-Madison , Madison, Wisconsin.

Abstract

Inflammation is involved in many prostate pathologies including infection, benign prostatic hyperplasia, and prostate cancer. Preclinical models are critical to our understanding of disease mechanisms, yet few models are genetically tractable. Here, we present a comparative quantitative proteomic analysis of urine from mice with and without prostate-specific inflammation induced by conditional prostate epithelial IL-1β expression. Relative quantification and sample multiplexing was achieved using custom 4-plex N,N-dimethyl leucine (DiLeu) isobaric tags and nanoflow ultrahigh-performance liquid chromatography coupled to high-resolution tandem mass spectrometry. Each set of 4-plex DiLeu reagents allows four urine samples to be analyzed simultaneously, providing high-throughput and accurate quantification of urinary proteins. Proteins involved in the acute phase response, including haptoglobin, inter-α-trypsin inhibitor, and α1-antitrypsin 1-1, were differentially represented in the urine of mice with prostate inflammation. Mass spectrometry-based quantitative urinary proteomics represents a promising bioanalytical strategy for biomarker discovery and the elucidation of molecular mechanisms in urological research.

KEYWORDS:

benign prostatic hyperplasia; inflammation; interleukin-1β; lower urinary tract symptoms, mass spectrometry; urine proteomics

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