RESUMO
Purified plant protein, Trichosanthin (TCS), was partially digested with chymotrypsin to produce peptide fragments. Two large fragments, I and II, have been isolated and identified as C-terminal peptides located at sequence positions between 79-234 and 107-234. Their molecular weights determined by electrophoretic mobility on SDS gel were 16,000 and 14,000 daltons. Studies using fluorescence quenching measurements by titrating anti-TCS Fab with TCS and fragment I and II showed that four epitopes were present in intact TCS and one each epitope in fragment I and II. One of the epitopes is therefore located between sequence positions 107 and 234.
Assuntos
Abortivos não Esteroides/imunologia , Abortivos/imunologia , Epitopos/análise , Fragmentos de Peptídeos/imunologia , Proteínas de Plantas/imunologia , Sequência de Aminoácidos , Animais , Dados de Sequência Molecular , Coelhos , TricosantinaAssuntos
Abortivos não Esteroides/imunologia , Abortivos/imunologia , Imunoglobulina E/imunologia , Idiótipos de Imunoglobulinas/imunologia , Proteínas de Plantas/imunologia , Animais , Anticorpos Monoclonais/imunologia , Ligação Competitiva , Reações Cruzadas , Epitopos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , TricosantinaRESUMO
An immunosuppressive protein was isolated from Trichosanthes kirilowii root tubers by a procedure involving acetone fractionation and ion exchange chromatography on CM-Sepharose. Homogeneity of the protein was demonstrated in immunoelectrophoresis, SDS-polyacrylamide gel electrophoresis, gel filtration, high performance liquid chromatography and a single NH2-terminal sequence. The protein had a molecular weight of 26,000, aspartic acid as the NH2-terminal amino acid and no cysteine or carbohydrates in its molecule. It inhibited ConA-induced transformation in lymphocytes isolated from spleens of CBA mice. The protein was also potent in inducing mid-term abortion in mice.