"Inject-mix-react-separate-and-quantitate" (IMReSQ) method for screening enzyme inhibitors.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Drug Discovery

Background:

• Regulatory enzymes are crucial therapeutic targets, with inhibitors holding potential as drug candidates.
• Identifying effective enzyme inhibitors often involves screening large combinatorial libraries.
• Protein farnesyltransferase from Entamoeba histolytica is a significant antiparasitic drug target.

Purpose of the Study:

• To introduce and validate a novel, low-volume method for screening enzyme inhibitors.
• To demonstrate the applicability of the method to a specific therapeutic target, protein farnesyltransferase.
• To identify new inhibitors for Entamoeba histolytica protein farnesyltransferase.

Main Methods:

• Developed the inject-mix-react-separate-and-quantitate (IMReSQ) method using nanoliter volumes.
• Utilized a capillary microreactor for controlled mixing and reaction of substrates, inhibitors, and enzymes.
• Employed capillary electrophoresis for separation and quantitation of reaction products and substrates.

Main Results:

• Successfully applied IMReSQ to screen inhibitors for Entamoeba histolytica protein farnesyltransferase.
• Identified three previously unknown inhibitors of the target enzyme.
• Demonstrated the quantitative capability of IMReSQ for ranking inhibitor potencies.

Conclusions:

• The IMReSQ method offers a highly efficient, low-volume approach for enzyme inhibitor screening.
• IMReSQ is effective for studying enzymes like protein farnesyltransferase, relevant to antiparasitic drug development.
• This method facilitates the identification and characterization of potential therapeutic compounds.