2143 QUANTITATIVE PROTEOMICS IDENTIFIES CERULOPLASMIN AS A POTENTIAL PROMOTER OF UROLITHIASIS

Abstract

You have accessJournal of UrologyStone Disease: Basic Research1 Apr 20112143 QUANTITATIVE PROTEOMICS IDENTIFIES CERULOPLASMIN AS A POTENTIAL PROMOTER OF UROLITHIASIS Cynthia Wright, Sarah Howles, David Trudgian, Benedikt Kessler, John Reynard, Jeremy Noble, Freddie Hamdy, and Benjamin Turney Cynthia WrightCynthia Wright Oxford, United Kingdom More articles by this author , Sarah HowlesSarah Howles Oxford, United Kingdom More articles by this author , David TrudgianDavid Trudgian Oxford, United Kingdom More articles by this author , Benedikt KesslerBenedikt Kessler Oxford, United Kingdom More articles by this author , John ReynardJohn Reynard Oxford, United Kingdom More articles by this author , Jeremy NobleJeremy Noble Oxford, United Kingdom More articles by this author , Freddie HamdyFreddie Hamdy Oxford, United Kingdom More articles by this author , and Benjamin TurneyBenjamin Turney Oxford, United Kingdom More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.2354AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Urinary proteins have been implicated as regulators of urolithiasis. Using label-free quantitative proteomics we identified several proteins that are more than 2-fold differentially abundant between pooled urine samples from stone formers (SF) and non stone formers (NSF). Our objective was to identify, validate and test the function of candidate proteins that may influence urolithiasis. METHODS Individual urine samples were normalised for natural variation in concentration using a total protein to creatinine ratio to calculate the volume contributing to the pooled sample. Differentially abundant proteins in pooled urine samples were identified using label-free nano-UPLC MSE analysis from two independent experiments using a total of 57 SF and 57 NSF. Enzyme-linked immunosorbent assay (ELISA) was used to validate the mass spectrometry findings and a calcium oxalate crystallization assay was performed to evaluate effects in vitro. RESULTS The urinary proteomic analysis identified 1063 proteins, of which 367 were unique to the stone former groups, 408 proteins were unique to the control pools and 288 proteins were identified for comparative quantification. Proteins found to be unique in stone-formers were involved in carbohydrate metabolism pathways and associated with disease states. 34 proteins demonstrated a consistent >2-fold change between stone formers and controls. For ceruloplasmin, one of the proteins shown to be more than two-fold up-regulated in the stone-formers, this observation was validated in individuals by ELISA. Moreover, in vitro crystallization assays demonstrated ceruloplasmin had a dose-dependent increase on calcium oxalate crystal formation. CONCLUSIONS This work has developed a label-free proteomics platform which may be utilised to identify biologically important urinary proteins in other urological diseases. We identified several urinary proteins that may influence urolithiasis. Ceruloplasmin, one such protein, was independently shown to be at higher levels in the urine of stone formers and to have a positive effect on calcium oxalate crystallization in vitro. Ceruloplasmin and the other proteins identified may play important roles in the biology of stone formation. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e858 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Cynthia Wright Oxford, United Kingdom More articles by this author Sarah Howles Oxford, United Kingdom More articles by this author David Trudgian Oxford, United Kingdom More articles by this author Benedikt Kessler Oxford, United Kingdom More articles by this author John Reynard Oxford, United Kingdom More articles by this author Jeremy Noble Oxford, United Kingdom More articles by this author Freddie Hamdy Oxford, United Kingdom More articles by this author Benjamin Turney Oxford, United Kingdom More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...