Johns Hopkins University

Memorial Sloan Kettering Cancer Center

Christopher D Ferris

Solomon H Snyder

Samie R Jaffrey

Paul J Tempst

Hediye Erdjument-Bromage

Soloman H Snyder

Jean-Pierre Mothet

Roscoe Brady, Jr

Herman Wolosker

Crystal C Watkins

Kevin Sheth

Takahashi Masaaki

Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. A method to detect proteins which contain nitrosothiols involves several steps. Nitrosylated cysteines are converted to tagged cysteines. Tagged proteins can then be detected, for example, by immunoblotting and/or can be purified by affinity chromatography. The method is applicable to the detection of S-nitrosylated proteins in cell lysates following in vitro S-nitrosylation, as well as to the detection of endogenous S-nitrosothiols in selected protein substrates.

Method for assaying protein nitrosylation

High levels of D-serine occur in mammalian brain, where it appears to be an endogenous ligand of the 'glycine site' of NMDA receptors. We have purified from rat brain a soluble enzyme that catalyzes the direct racemization of L-serine to D-serine. Purified serine racemase has a molecular weight of 37 kDa and requires pyridoxal 5'-phosphate for its activity. The enzyme is highly selective toward L-serine, failing to racemize any other amino acid tested. We have also identified polynucleotide sequences that encode mammalian, including human, serine racemase. Compounds that modulate the activity of mammalian serine racemase are useful for treating conditions and diseases that involve overstimulation of NMDA receptors, such as stroke and various neurodegenerative diseases.

Mammalian serine racemase

Disclosed are methods for preventing or treating a gastrointestinal (GI) disorder in a mammal such as a human patient. In one embodiment, the methods include administering to the mammal a therapeutically effective amount of a compound that modulates a nitric oxide (NO) signaling pathway, particularly in GI neurons. Methods of the invention are particularly useful for the treatment (including prophylactic treatment) of diabetic gastropathies and other GI disorders.

Methods for prevention and treatment of gastrointestinal disorders

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Franklin, TN, United States of America

Christopher D Ferris