Compound heterozygosis in AADC deficiency: A complex phenotype dissected through comparison among heterodimeric and homodimeric AADC proteins

Compound heterozygosis is the most diffuse and hardly to tackle condition in aromatic amino acid decarboxylase(AADC) deficiency, a genetic disease leading to severe neurological impairment. Here, by using an appropriatevector, we succeeded in obtaining high yields of AADC protein and characterizing two new heterodimers,T69M/S147R and C281W/M362T, detected in two AADC deficiency patients. We performed an extensive bio-chemical characterization of the heterodimeric recombinant proteins and of the related homodimers, by a com-bination of dichroic andfluorescence spectroscopy and activity assays together with bioinformatic analyses. Wefound that T69M/S147R exhibits negative complementation in terms of activity but it is more stable than the av-erage of the homodimeric counterparts. The heterodimer C281W/M362T retains a nearly good catalytic effi-ciency, whereas M362T homodimer is less affected and C281W homodimer is recovered as insoluble. Theseresults, which are consistent with the related phenotypes, and the data emerging from previous studies, suggestthat the severity of AADC deficiency is not directly explained by positive or negative complementation phenom-ena, but rather depends on: i) the integrity of one or both active sites; ii) the structural and functional propertiesof the entire pool of AADC proteins expressed. Overall, this integrated and cross-sectional approach enablesproper characterization and depicts the functional result of subunit interactions in the dimeric structure andwill help to elucidate the physio-pathological mechanisms in AADC deficiency.