A refined quaternary structure of Androctonus australis Hemocyanin

The quaternary structure of the (4 X 6)-mer hemocyanin from the Scorpion Androctonus australis previously published [Lamy, J., Bijlholt, M. M. C., Sizaret, P.-Y., Lamy, J., and van Bruggen, E. F. J. (1981) Biochemistry, 20, 1849-1856] has been refined. The relative positions in the half molecule of subunits Aa 3A and Aa 3B compared to those of Aa 3C and Aa 5B have been established by double labeling of the (2 X 6)-mer with binary mixtures of subunit-specific Fab fragments. The results show that subunits Aa 3B and Aa 5B are located in the same hexamer while Aa 3A and Aa 3C are in the other half of the (2 X 6)-mer. The choice of the enantiomer was deduced from a careful examination of electron micrographs of the native molecule. Finally a position was assigned to each of the 24 subunits on the flip and flop faces as defined by Van Heel and Frank [Ultramicroscopy, 6, 187-194 (1981)].