Abstract
A rapid and sensitive assay has been developed for the factor-dependent dissociation of eukaryotic ribosomes. This assay takes advantage of the observation that initiation factor eIF-2 will bind Met-tRNAf met to 40 S subunits but not to 80 S ribosomes. Incubation of wheat germ ribosomes at 1 mm Mg2+ results in their dissociation into 40 S subunits. These subunits spontaneously reassociate when the Mg2+ concentration is raised to 4 mm. However, if the incubation at 1 mm Mg2+ is carried out in the presence of an extract containing a ribosome dissociation factor, a certain portion of the subunits will fail to reassociate when the Mg2+ concentration is raised to 4 mm. The 40 S subunits remaining due to the presence of the dissociation factor can bind [35S]Met-tRNAf met in the presence of wheat germ eIF-2. The [35S]Met-tRNAf met bound to the 40 S subunits is readily detected by its retention on a Millipore filter.
Original language | English (US) |
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Pages (from-to) | 238-243 |
Number of pages | 6 |
Journal | Analytical biochemistry |
Volume | 93 |
Issue number | C |
DOIs | |
State | Published - 1979 |
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology