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JoVE Journal Biochemistry
Nitropeptide Profiling and Identification Illustrated by Angiotensin II
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Nitropeptide Profiling and Identification Illustrated by Angiotensin II

Article DOI: 10.3791/59391-v • 07:31 min • June 16th, 2019
June 16th, 2019 •
1,2, 3, 2,4,5,6
1Mass Spectrometry Core Facility, School of Life Sciences, Westlake University, 2Department of Biological Sciences, University of Notre Dame, 3Department of Urology, Shanghai East Hospital, Tongji University School of Medicine, 4Boler-Parseghian Center for Rare and Neglected Diseases, University of Notre Dame, 5Harper Cancer Research Institute, University of Notre Dame, 6Tumor Microenvironment and Metastasis Program, Indiana University Melvin and Bren Simon Cancer Center

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Summary
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Proteomic profiling of tyrosine-nitrated proteins has been a challenging technique due to the low abundance of the 3-nitrotyrosine modification. Here we describe a novel approach for nitropeptide enrichment and profiling by using Angiotensin II as the model. This method can be extended for other in vitro or in vivo systems.

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Nitropeptide Profiling Nitropeptide Identification Chemical Approach Mass Spectrometry Enhanced Sensitivity In Vitro Biochemical Systems Endogenous Biological Tissues Nitro-angiotensin II Desalting Reverse-phase Solid-phase Extraction (SPE) Column Methanol Water Elution Primary Amine Alkylation Triethylammonium Bicarbonate Solution Formaldehyde Sodium Cyanoborohydride Labeling Reaction
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