Characterization and inactivation of an agmatine deiminase from Helicobacter pylori
| Autor: | Brian Knuckley, Paul R. Thompson, Corey P. Causey, Leslie L. Lovelace, Lukasz Lebioda, Justin E. Jones, Heather Flick |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Agmatine
Arginine Hydrolases Amidines Crystallography X-Ray Biochemistry Article Virulence factor Substrate Specificity chemistry.chemical_compound Drug Discovery Hydrolase Humans Amino Acid Sequence Molecular Biology chemistry.chemical_classification Binding Sites Innate immune system Helicobacter pylori Organic Chemistry Citrullination Recombinant Proteins Agmatine deiminase Kinetics Enzyme chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization |
| Zdroj: | Bioorganic Chemistry. 38:62-73 |
| ISSN: | 0045-2068 |
| Popis: | Helicobacter pylori encodes a potential virulence factor, agmatine deiminase (HpAgD), which catalyzes the conversion of agmatine to N-carbamoyl putrescine (NCP) and ammonia - agmatine is decarboxylated arginine. Agmatine is an endogenous human cell signaling molecule that triggers the innate immune response in humans. Unlike H. pylori, humans do not encode an AgD; it is hypothesized that inhibition of this enzyme would increase the levels of agmatine, and thereby enhance the innate immune response. Taken together, these facts suggest that HpAgD is a potential drug target. Herein we describe the optimized expression, isolation, and purification of HpAgD (10–30 mg/L media). The initial kinetic characterization of this enzyme has also been performed. Additionally, the crystal structure of wild-type HpAgD has been determined at 2.1 Å resolution. This structure provides a molecular basis for the preferential deimination of agmatine, and identifies Asp198 as a key residue responsible for agmatine recognition, which has been confirmed experimentally. Information gathered from these studies led to the development and characterization of a novel class of haloacetamidine based HpAgD inactivators. These compounds are the most potent AgD inhibitors ever described. |
| Databáze: | OpenAIRE |
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