Crystallographic study of the interaction of the anti-HIV lectin actinohivin with the α(1-2)mannobiose moiety of gp120 HMTG.

Abstract

Actinohivin (AH) is a new potent anti-HIV lectin of microbial origin. In order to modify it to produce a more efficient drug, its three-dimensional structure has previously been determined with and without the target α(1-2)mannobiose moiety of the high-mannose-type glycan (HMTG) attached to HIV-1 gp120. However, ambiguity remained in the structures owing to packing disorder that was possibly associated with peptide fragments attached at the N-terminus. To resolve these problems, the duration of cultivation of the AH-producing strain was examined and it was found that in a sample obtained from a 20 d culture the heterogeneous fragments were completely removed to produce mature AH with high homogeneity. In addition, the purification procedures were simplified in order to increase the yield of AH and the addition of solvents was also examined in order to increase the solubility of AH. AH thus obtained was successfully crystallized with high reproducibility in a different form to the previously obtained crystals. The crystal diffracted well to beyond 1.90 Å resolution and the crystallographic data suggested that it contained no packing disorder.