The interactions between the N-terminal and C-terminal domains of the human UDP-glucuronosyltransferases are partly isoform-specific, and may involve both monomers.

The pathological mutation Y486D was previously shown to reduce the activities of the UDP-glucuronosyltransferases (UGTs) 1A1 and 1A6 by about 88% and 99%, respectively. Surprisingly, the corresponding mutation in UGT1A9 (Y483D) doubled the Vmax of scopoletin glucuronidation, whereas the entacapone glucuronidation rate was decreased by about 50%. Due to the primary structure identity of the C-terminal half of all the human UGTs of the 1A subfamily, the sharp differences between them in the effect of a mutation deep inside the C-terminal half suggested that there are isoform-specific interactions between the variable N- and the conserved C-terminal halves. In dimeric enzymes, like the UGTs, such interactions might either occur within the same polypeptide, or between opposite monomers. The latter implies functional monomer-monomer interactions, and this was investigated using hetero-dimeric UGTs. Insect cells were co-infected with mixtures containing different combinations of recombinant baculoviruses encoding either UGT1A4 or 1A9Sol. The UGT1A4 was selected because it glucuronidates neither entacapone nor scopoletin at significant rates. The active enzyme in these hetero-dimers was 1A9Sol, a truncation mutant of UGT1A9 that exhibited a very low ratio of entacapone to scopoletin glucuronidation rates. Interestingly, the ratio of entacapone to scopoletin glucuronidation rates in the co-infected cells was dependent on, and markedly increased with, the probability that 1A9Sol forms hetero-dimers with UGT1A4. In addition, the apparent Km for entacapone in the hetero-dimers was much lower than in 1A9Sol, and resembled the corresponding value in full-length UGT1A9. The results, thus, revealed important monomer-monomer interactions within the UGTs.