Eukaryotic mRNA Cap Binding Protein: Purification by Affinity Chromatography on Sepharose-Coupled m7GDP
Abstract
A 24,000-dalton polypeptide that binds strongly and can be specifically crosslinked to the 5'-terminal cap structure m7GpppN in eukaryotic mRNAs has been detected in protein synthesis initiation factor preparations [Proc. Natl. Acad. Sci. USA (1978) 75, 4843--4847]. This polypeptide has been purified to apparent homogeneity by one chromatographic passage through an affinity resin prepared by coupling the levulinic acid O2',3'-acetal of m7GDP to AH-Sepharose 4B. Translation, in HeLa cell extracts, of capped mRNAs including Sindbis virus, reovirus, and rabbit globin mRNAs was stimulated by the cap-binding protein under conditions that did not increase translation of noncapped RNAs of encephalomyocarditis virus and satellite tobacco necrosis virus.
- Publication:
-
Proceedings of the National Academy of Science
- Pub Date:
- September 1979
- DOI:
- 10.1073/pnas.76.9.4345
- Bibcode:
- 1979PNAS...76.4345S