Antigenic determination fragments of alpha-momorcharin.

Abstract Alpha momorcharin is a protein isolated from the bitter gourd. It has a number of biological activities including induction of abortion, inhibition of tumor growth and anti-HIV. All these activities may be related to the ribosome-inhibiting activity of the protein. Repeated use of αMMC can elicit an antigenic response which may neutralize its biological activity. To overcome this problem, we need to know which part of the molecule is the antigenic determinant. In this study, we constructed a random fragment expression library from the αMMC cDNA and screened it with three anti-αMMC sera. A total of 9 positive clones were picked and sequenced. Based on the sequence information obtained, we were able to deduce three regions at which antibodies raised against native αMMC seem to interact. These regions are residues 1–14, residues 71–136 and residues 195–222. Mapping of these regions against a 3D model of αMMC indicates that they all are located on the surface of the molecule. As residues 71–136 are found to be in close proximity to the active site involved in ribosome inactivation, treatment with a monoclonal antibody directed to this area was shown to be effective in inactivating the inhibitory effect of αMMC on in vitro protein synthesis.