The Purification of Monoamine Oxidase Type A from Beef Brain Mitochondria
Siegel, David Michael
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The purification of Monoamine Oxidase Type A, MAO A, an enzyme involved in the oxidation of amines to aldehydes, was performed for the purpose of comparison between its isoenzyme, MAO B. The enzyme, which is blocked by the intake of many anti-depressant drugs, was performed from beef brain mitochondria, so an interspecies comparison could be done. The mitochondria were prepared from the beef brain by a series of centrifugations. The primary step was the extraction of the enzyme from the outer mitochondrial membrane. This was performed with five different detergents to observe which was most effective in the enzyme extraction. This same process was performed again; however, with the addition of 1M KCI, to observe if changing the "ionic strength" of the solution would solubilize more protein away with the membrane. The detergent was then removed by dialysis. The first purification step was treating the MAO A solution with Trypsin covalently bound to beads. This was done to inspect its effect on the protein in an attempt to leave the MAO A in solution with fewer proteins, and closer to its pure state.