Metabolic Control Analysis for Drug Target Prioritization in Trypanosomatids

Area of Science:

• Parasitology
• Biochemistry
• Systems Biology

Background:

• Enzyme essentiality via gene knockout/knockdown is insufficient for prioritizing therapeutic targets in trypanosomatids.
• Most analyzed enzymes in these parasites are essential, necessitating additional criteria for drug target selection.

Purpose of the Study:

• To introduce Metabolic Control Analysis (MCA) as a method for prioritizing enzymes as drug targets in trypanosomatid metabolic pathways.
• To explain how MCA quantifies enzyme control over pathway flux using flux control coefficients ([Formula: see text]).

Main Methods:

• Review of three approaches to determine flux control coefficients: in vitro pathway reconstitution, manipulation of enzyme activities, and in silico kinetic modeling.
• Emphasis on the need for accurate experimental data (enzyme activities, metabolite concentrations, pathway fluxes) for MCA.
• Illustration of MCA methodology using trypanothione metabolism in *Trypanosoma cruzi*.

Main Results:

• MCA establishes the distribution of metabolic control, identifying key flux-controlling steps within a pathway.
• This allows for a metabolic-based ranking of pathway enzymes as potential drug targets.

Conclusions:

• Metabolic Control Analysis provides a robust framework for prioritizing drug targets in trypanosomatids by quantifying enzyme control.
• The described MCA strategy and protocols are applicable to any metabolic pathway in these parasites.