Hi,
I would like to know the best method to check the interaction between a proteine (cruzipain, cysteine protease) and a drug. It has been purposed to our project to evaluate protein-drug interaction with EliSA test (fixing drug or protein following our capabilities) but recent data may suggest to think about chromatography ( HPLC ? HPAC ?) or even LCMS–MS. I would like to have suggestions or advice in this subject. Thanks
Cordially
Hi,
What is it you specifically would like to know? Qualitative binding, stochiometry, binding strength? Or something else? This will determine what method can be used best. Are molecules interacting covalently, or non-covalently?
Electrospraying a mixture of the protein and drug in solution into the mass spectrometer under native (i.e. binding) conditions will enable you to look at some aspects of their interactions.
Hope this helps.
Hi Eef Dirksen
First of all, thank you for you answer. My purpose is to check or determine the molecular target (protein or RNA) of common drugs such as atovaquone, doxycyline, azythromyicin (for example) in differents pathogens coming from kinetoplastidae or apicomplexa phylum where data are not documented.
We would like to carry out experiments to know qualitative** binding between the drug and the different proteins (from a parasite lysate). Drugs used in our research are antibiotics, they were previously documented with non covalent interaction (Inhibitor in a Michaelis Mantenn system).
Our second hypothesis was to think drug targets the same protein in 2 different parasites, so we would use a matrix with this protein to fix our drug.
Cordially
Hi
I suggest to use Electrospray ionization (ESI) is the Most frequently used ionization methods for the analysis of this molecule Or MALDI/TOF/TOF
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