Molecular properties of aminopeptidase Ey as a zinc-metalloenzyme.

1. Aminopeptidase Ey from hen's egg yolk contains 1.0 g atom of zinc/mol of a subunit having molecular weight of 150 kDa. The inactive, Zn(2+)-free apoenzyme was reactivated by Co2+, Mn2+, Ca2+, Cd2+, Cu2+ and Ni2+ in addition to Zn2+, whereas Mg2+ and Fe2+ were ineffective. 2. The enzymatical properties of reconstituted enzymes, except for Zn(2+)-reconstituted enzyme, differed from native enzyme. The values for the activation energy were calculated by aminopeptidase Ey and Co(2+)-reconstituted enzyme. 3. The isoelectric point of the enzyme was about 2.8 as determined by isoelectric focusing. An asialo form of the enzyme, obtained by treatment with Arthrobacter sialidase, had an isoelectric point of 4.4. 4. The amino terminal sequence of aminopeptidase Ey was determined to be acyl-Xaa-Xaa-Pro-Glu-Ala-Ala-Ser-Leu-Pro-Gly. There was no identity with any known sequences of aminopeptidase.