The University of Texas System

Emil Martin

Ferid Murad

Andrei Nedospasov

Nataliya Beda

Tatiana Pimenova

Methods and compositions are disclosed for quantifying and/or identifying a nitroderivative compound, or a residue thereof, and are applicable for assessing in vivo nitrative stress and for aiding in diagnosis of mammalian pathologies such as asthma, atherosclerosis, Alzheimer's disease, inflammation, ischemia, Parkinson's disease, and cancer. A preferred method comprises a) providing a sample containing the nitroderivative compound, or residue thereof, in the form of a diazo compound or residue thereof; b) azo-coupling the diazo compound, or residue thereof, to a target compound capable of producing a signal, to yield an azo-coupled compound; c) isolating the nitroderivative compound, or residue thereof, or the azo-coupled compound; d) quantitating and/or detecting the signal, to yield a quantitation and/or detection result; and e) determining from the quantitation and/or detection result the identity and/or concentration of the nitroderivative compound, or residue thereof, in the sample.

Methods of detecting nitrotyrosine and aminotyrosine residues of peptides and proteins

A method of screening a substance of interest for heme independent modulation of enzymatic activity of soluble guanylyl cyclase (sGC) is disclosed, comprising (a) obtaining &#x3b1;&#x3b2;mutant sGC enzyme; (b) determining activity of the mutant enzyme for forming cGMP from GTP in the presence of the substance of interest in a reaction medium; (c) determining activity as in step (b), except in the absence of the substance of interest; optionally, (d) including an activator other than the substance of interest in steps b) and c); e) comparing results of (b)&#x2013;(d) to yield a comparison result; and f) from that value of that result, assessing activity of the substance of interest for modulating cGMP production by the mutant enzyme. Increased or decreased formation of cGMP in the presence of the substance of interest indicates activity of the substance for modulating heme independent cGMP production.

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Houston, TX, United States of America

Emil Martin