| Original language | English (US) |
|---|---|
| Pages (from-to) | 256-265 |
| Number of pages | 10 |
| Journal | Methods in Enzymology |
| Volume | 60 |
| Issue number | C |
| DOIs | |
| State | Published - Jan 1 1979 |
| Externally published | Yes |
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry
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In: Methods in Enzymology, Vol. 60, No. C, 01.01.1979, p. 256-265.
Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - Millipore Filtration Assay for AUG-Directed Met-tRNAf Binding to 40 S and 80 S Ribosomes
AU - Chatterjee, Bandana
AU - Dasgupta, Asim
AU - Majumdar, Alokes
AU - Palmieri, Steven
AU - Gupta, Naba K.
N1 - Funding Information: Two-dimensional gel electrophoresis is a powerful analytical technique, widely used for separating complex mixtures of proteins. The method developed by O'Farrell" uses isoelectrofocusing in the presence of urea in the first dimension and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis in the second dimension; it is preferred to other methods because proteins can be analyzed in their native state by simply omitting urea in the isoelectric focusing gel. We have slightly modified the O'Farrell procedure in order to investigate the subunit interaction and function of eukaryotic initiation factor 2 (eIF-2). eIF-2 appears to be required in the first step of initiation of protein Investigations from the authors' laboratoryw ere supported by U.S. Public Health Service Research Grant AM 18477. P. H. O'Farrell, J. Biol. Chem. 250, 4007 (1975). Funding Information: i This work was supported by NIH Research Grants GM-18796 and GM-22079. z The nomenclature for the eukaryotic peptide chain initiation factors used in this chapter and in chapters \[4\]a nd \[5\]a re: ElF-I, Met-tRNAr binding factor; TDF (also termed EIF-2), ternary complex (Met-tRNAr. ElF-1 .GTP) dissociation factor; Co-ElF-1, EIF-I stimulat-ory protein; EIF-3, ribosomal subunit joining factor; DF, 80 S ribosome dissociation factor. According to the Anderson et al. proposed nomenclature \[W. F. Anderson, L. Bosch, W. E. Cohn, H. Lodish, W. C. Merrick, H. Weissbach, H. G. Wittmann, and I. G. WooI,FEBS Lett. 76, 1( 1977)\],E IF-1 is elF-2 and EIF-3 is elF-5. The identities of the~other factor~ are not clear. :3 y. C. Chen, C. L. Woodley. K. K. Bose, and N. K. Gupta, Biochem. Biophys. Res. Commun. 48, I (1972). 4 N. K. Gupta, C. L. Woodley, Y. C. Chen, and K. K. Bose, J. Biol. Chem. 248, 4500 (1973). G. L. Dettman and W. M. Stanley, Biochim. Biophys. Acta 287, 124 (1972). 6 D. H. Levin, D. Kyner, and G. Acs, Proc. Natl. Acad. Sci. U.S.A. 70, 41 (1973). 7 M. H. Schreier, and T. Staehelin, Nature (London), New Biol. 242, 35 (1973). 8 R. S. Ranu, and I. G. Wool, Nature (London) 257, 616 (1975). 9 B. Safer, W. F. Anderson, and W. C. Merrick, J. Biol. Chem. 250, 9067 (1975). 1o W. Filipowicz, J. M. Sierra, and S. Ochoa, Proc. Natl. Acad. Sci. U.S.A. 72, 3947 (1975). H B. Chatterjee, A. Dasgupta, S. Palmieri, and N. K. Gupta,J. Biol. Chem. 251, 6379 (1976). Jz A. Majumdar, R. Roy, A. Das, A. Dasgupta, and N. K. Gupta, Biochem. Biophys. Res. Commun. 78, 161 (1977).
PY - 1979/1/1
Y1 - 1979/1/1
UR - http://www.scopus.com/inward/record.url?scp=0018401887&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0018401887&partnerID=8YFLogxK
U2 - 10.1016/S0076-6879(79)60023-X
DO - 10.1016/S0076-6879(79)60023-X
M3 - Article
C2 - 459901
AN - SCOPUS:0018401887
SN - 0076-6879
VL - 60
SP - 256
EP - 265
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -