A Method for Monitoring Protein S-Nitrosylation

S-nitrosylation is the addition of a nitric oxide group to a cysteine group in a protein sequence to form an S-nitrosothiol derivative. JHU scientists developed a method in which a test sample comprising at least one protein substrate is treated with an alkylthiolating agent to block free thiol groups on the substrate. Nitrosothiol bonds on the protein substrate are reduced to form free thiol groups. The free thiol groups on the protein are reacted with a tagged, activated mixed disulfide which can be detected and measured. JHU scientists have detected proteins sensitive to both exogenous and endogenous S-nitrosylation using normal and mutant rodent brain lysates. Description (Set) Johns Hopkins University is seeking licensees for a simple assay to detect S-nitrosylation of neuronal proteins. A hallmark of neurological diseases such as ALS, Alzheimer�s or Parkinson�s disease is the accumulation of aggregated proteins that interfere with normal nerve cell activity. While many neurodegenerative diseases mechanisms are unknown, alterations in nitrosative/oxidative stress may be partially responsible for pathological protein aggregation. Nitrosative stress results from high intracellular nitric oxide (NO) levels. NO mediates S-nitrosylation, which is a post-translational modification of a polypeptide sequence that can alter protein structure and activity. To date, multiple proteins associated with neurodegenerative disease were found to be S-nitrosylated (for review, see Nakamura 2008 and Chung 2006). Detection of S-nitrosylation in proteins is a recognized challenge. JHU scientists have determined a simple, sensitive and specific method for detection of protein S-nitrosylation. Identified S-nitrosylated proteins from neuronal tissue are potential therapeutic targets for neurodegenerative diseases.
• S-nitrosylation assay can be performed without purifying individual protein substrates from tissue to expand applicability for measurement of nitrosylated proteins both in vitro and in vivo.
• S-nitrosylation assay can be performed without using photolytic- chemiluminescent detection for a specific and simplified assay. Proposed Use (Set) This technology can be commercialized as a detection kit for measuring protein nitrosylation. This technology can also be commercialized as a method to identify agents that modulate protein S-nitrosylation which may be useful for therapeutic drug development Patent (Set) US Patent No. 6,806,057; Int'l Pub. WO 02/39119; US Patent No. 7,001,7.8;

US 6,806,057
US 700,178

Inventor(s): Snyder, Solomon H.

Type of Offer: Licensing

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