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

Author:

Hao Ling1,Thomas Samuel2,Greer Tyler3,Vezina Chad M.245,Bajpai Sagar6,Ashok Arya6,De Marzo Angelo M.7,Bieberich Charles J.68,Li Lingjun123,Ricke William A.1259ORCID

Affiliation:

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.

Funder

HHS | NIH | National Institute of Environmental Health Sciences (NIEHS)

HHS | NIH | National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)

HHS | National Institutes of Health (NIH)

Publisher

American Physiological Society

Subject

Physiology

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