Antigenic determination fragments of alpha-momorcharin.
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 alphaMMC 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 alphaMMC cDNA and screened it with three anti-alphaMMC 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 alphaMMC 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 alphaMMC 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 alphaMMC on in vitro protein synthesis.
Leung
K C
KC
Department of Biochemistry, Chinese University of Hong Kong, Shatin, N.T.
Meng
Z Q
ZQ
Ho
W K
WK
D000020:Abortifacient Agents, Nonsteroidal; D019380:Anti-HIV Agents; D000911:Antibodies, Monoclonal; D000972:Antineoplastic Agents, Phytogenic; D001665:Binding Sites; D017931:DNA Primers; D000939:Epitopes / Q000032:analysis* / Q000737:chemistry; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000276:immunology; D010940:Plant Proteins / Q000737:chemistry* / Q000276:immunology* / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D012269:Ribosomal Proteins; D054788:Ribosome Inactivating Proteins; D012270:Ribosomes / Q000187:drug effects
D000020:Abortifacient Agents, Nonsteroidal; D019380:Anti-HIV Agents; D000911:Antibodies, Monoclonal; D000972:Antineoplastic Agents, Phytogenic; D017931:DNA Primers; D000939:Epitopes; D010446:Peptide Fragments; D010940:Plant Proteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C039582:MMC protein, Momordica charantia; D054788:Ribosome Inactivating Proteins