Mammalian polyamine oxidase (PAO) is a member of the monoamine oxidase family of flavoproteins and catalyzes the oxidation of polyamines. Polyamines are essential for cell growth and proliferation, although their specific role in the cell is still a matter of discussion. A crystal structure of the enzyme will help us determine basic chemical and structural differences among PAO’s and the specific roles of polyamine metabolites.<br/>Currently, there are crystal structures of the yeast and maize PAO enzymes available, but these enzymes are only 20% identical to the mammalian PAOs. Recent attempts to crystallize mouse polyamine oxidase (mPAO) have been unsuccessful. The identity between the human and mouse PAO is greater than 80%. Therefore, in order to continue our pursuit of obtaining the first crystal structure of a mammalian PAO we have decided to crystallize the human PAO (hPAO).<br/>In order to crystallize hPAO we have been able to express and purify the hPAO protein with a histidine tag. Enzyme activity assays where performed, showing that hPAO catalyzes the oxidation of polyamines. Furthermore, a sample of the purified enzyme has been submitted to the X-Ray Crystallization Laboratory Core at University of Texas Health Science Center at San Antonio for crystallization trials.

Expression and purification of human polyamine oxidase

251st National Meeting (2016)

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