Monoclonal antibody specific for yeast elongation factor 3
- 19 June 1984
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 23 (13), 3055-3063
- https://doi.org/10.1021/bi00308a032
Abstract
Hybridomas were prepared by fusing mouse myeloma (P3 .times. 63 Ag8) cells of mice immunized with a yeast fraction enriched with respect to nonribosomal translational components. Cloned hybridoma lines were grown in the form of ascites tumors, and the monoclonal antibodies produced were purified from the ascites fluid by chromatography on DEAE-Affi-Gel Blue. One of the antibodies, from a hybridoma cell line designated as PSH-1, inhibited the translation of natural mRNA and poly(U) and polysomal chain elongation in a cell-free protein-synthesizing system from yeast. Resolution and partial purification of the elongation factors [EF] indicated that the monoclonal antibody from PSH-1 did not interact with EF-1 or EF-2 but reacted with and inactivated EF-3, the 125,000 MW additional elongation factor specifically required with yeast ribosomes. The EF-3 purified from the cytosol by immunoaffinity chromatography was comparable to that prepared by ion-exchange chromatography. Evidently, EF-3 was essential for the translation of natural mRNA as well as poly(U), was associated with polysomes but not ribosomal subunits, and was required for every cycle in the elongation phase of protein synthesis.Keywords
This publication has 21 references indexed in Scilit:
- Magnesium-dependent interaction of 30S ribosomal subunits with antibodies to N6,N6-dimethyladenosineBiochemistry, 1980
- The effect of antibodies against Escherichia coli small ribosomal subunit proteins on protein synthesis by rat liver ribosomes.Journal of Biological Chemistry, 1980
- the acidic ribosomal phosphoproteins from Saccharomyces cerevisiaeBiochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1980
- Protein synthesis in rabbit reticulocytes. Demonstration of the requirements for eIF-2 and Co-eIF-2A for peptide chain initiation using immune sera.Journal of Biological Chemistry, 1979
- Function of individual E. coli 30 S ribosomal proteins as determined by in situ immunospecific neutralization : a tentative classificationBiochimie, 1979
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- The preparation and characterization of a cell-free system from Saccharomyces cerevisiae that translates natural messenger ribonucleic acid.Journal of Biological Chemistry, 1979
- Regulation of protein synthesis in reticulocyte lysates: immune serum inhibits heme-regulated protein kinase activity and differentiates heme-regulated protein kinase from double-stranded RNA-induced protein kinase.Proceedings of the National Academy of Sciences, 1979
- Determination of protein synthesis initiation factor levels in crude lysates of Escherichia coli by a sensitive radioimmune assayArchives of Biochemistry and Biophysics, 1978
- Dissimilarity in protein chain elongation factor requirements between yeast and rat liver ribosomes.Journal of Biological Chemistry, 1977