Monoclonal antibodies to elongation factor-1? inhibit in vitro translation in lysates of Sf21 cells
| Autor: | N. R. Chamberlain, Melissa K. Stuart |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Physiology
medicine.drug_class Immunoprecipitation Blotting Western Spodoptera Monoclonal antibody Biochemistry Cell Line Mice Peptide Elongation Factor 1 medicine Protein biosynthesis Animals Humans Sf21 Protein Synthesis Inhibitors Chromatography Mice Inbred BALB C biology Antibodies Monoclonal General Medicine biology.organism_classification Precipitin Tests Molecular biology In vitro Cell culture Protein Biosynthesis Insect Science biology.protein Electrophoresis Polyacrylamide Gel Female Antibody |
| Zdroj: | Archives of Insect Biochemistry and Physiology. 52:17-34 |
| ISSN: | 1520-6327 0739-4462 |
| DOI: | 10.1002/arch.10061 |
| Popis: | Elongation factor-1alpha (EF-1alpha) is an enzyme that is essential for protein synthesis. Although EF-1alpha offers an excellent target for the disruption of insect metabolism, agents known to interfere with EF-1alpha activity are toxic to humans. In this article, we describe the development of monoclonal antibodies (MAbs) that can disrupt the activity of insect EF-1alpha without cross-reacting with the human enzyme. MAbs were generated to EF-1alpha from Sf21 cells derived from the fall armyworm, Spodoptera frugiperda, by immunizing mice with EF-1alpha eluted from SDS-PAGE gels. The MAbs reacted with EF-1alpha in eggs and first through fifth instars of the fall armyworm in immunoblots of SDS-PAGE gels, but did not recognize EF-1alpha in human carcinoma cells and normal tissues. MAbs with the ability to recognize EF-1alpha in its native conformation, identified through immunoprecipitation experiments, were added to Sf21 cell lysates to determine whether the antibodies could inhibit incorporation of [(35)S]methionine into newly synthesized in vitro translation products. Of the four EF-1alpha-specific MAbs tested, three significantly inhibited protein synthesis when compared to the negative control antibody (P < 0.001, one-way ANOVA; followed by Dunnett's test, P < 0.05). |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text