Structure-based drug design with Kynurenine Aminotransferase-I

Summary

Human kynurenine amino transferase (KAT-1) forms the basis of a structure-based drug design study with implications for schizophrenia.

Supervisor(s)

Dr W Bret Church

Research Location

Faculty of Pharmacy

Program Type

Masters/PHD

Synopsis

Schizophrenia is a psychosis that affects 1% of the population world wide. The molecular mechanisms that trigger schizophrenia remain elusive. Several environmental risk factors have been considered, however studies performed with twins suggest that it is a highly heritable disease. Recently attention has shifted from dopamine and serotoninergic systems (dopaminergic hyperactivity) to the additional involvement of a glutamatergic neurotransmission system in schizophrenia.    One of the most widely distributed receptors at the central nervous system is the glutamatergic NMDA receptor(the NMDA glutamatergic hypofunction model). This receptor is highly sensitive to the levels of molecules such as glutamate and D-serine (the endogenous co-agonists of this receptor). Overactivity in Kynurenine Aminotransferase (KAT), which synthesizes kynurenic acid, an NMDA antagonist, may be an important link according to the glutamatergic hypofunction model of schizophrenia.  To pursue large scale production of KAT-1 E.coli will be used for expression, to obtain the protein required for crystallization studies.   Protein content can be estimated using a number of techniques and enzyme activity of the KAT-1 can be confirmed using kynurenine and pyruvate as substrates in HPLC assays.  The methods could use expression vectors with cleavable His6 tags that provide for the use of Ni-NTA nickel-chelating resin in purification but that can easily be removed if there appears to be any interference with the crystallization process.   Pure protein is required for crystallization, which allows determination of the crystallographic structure of the enzyme with inhibitors.  These outcomes complement the strategies for the rational design of more potent inhibitors of KAT.

Additional Information

Techniques used will be HPLC, X-ray crystallography, isothermal calorimetry, molecular biology, molecular modeling, protein purification

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Keywords

Schizophrenia, Crystallisation, protein, complex, structure, drug design, inhibitors, rational drug design, Brain & nervous system disorders, Therapeutics & adverse drug effects, Cell biology, Genes in biology & medicine, Pharmacology & therapeutics

Opportunity ID

The opportunity ID for this research opportunity is: 215

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