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. 1987 Jun 15;244(3):523–532. doi: 10.1042/bj2440523

A rapid and preparative method for the separation of yeast ribosomal proteins by using high-performance liquid chromatography.

G J Threadgill 1, R C Conrad 1, M Cannon 1, G R Craven 1
PMCID: PMC1148027  PMID: 3328611

Abstract

Ribosomal proteins from the yeast Saccharomyces cerevisiae were separated, on a preparative scale, by ion-exchange h.p.l.c. Proteins from the small and large ribosomal subunits were resolved, respectively, into 33 and 23 peaks, and most of the proteins present in these peaks were identified by using one- and two-dimensional gel electrophoresis. Several of the peaks appeared to contain a single protein uncontaminated by other species. Ribosomal proteins were also separated by using reverse-phase h.p.l.c. Analysis of the peaks resolved indicated that the order of elution for the proteins of both ribosomal subunits is, in certain cases, different for each of the two h.p.l.c. techniques used. Thus a combination of the two chromatographic methods employed here has the potential to facilitate the rapid and preparative separation of each of the proteins present in yeast ribosomes.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Capel M., Datta D., Nierras C. R., Craven G. R. Preparative ion-exchange high-performance liquid chromatography of bacterial ribosomal proteins. Anal Biochem. 1986 Oct;158(1):179–188. doi: 10.1016/0003-2697(86)90607-x. [DOI] [PubMed] [Google Scholar]
  2. Dalrymple P. N., Gupta S., Regnier F., Houston L. L. Separation of eucaryotic ribosomal proteins by high-performance liquid chromatography using a new cation-exchange resin and LiCl elution in 4 M urea. Biochim Biophys Acta. 1983 Jan 25;755(2):157–162. doi: 10.1016/0304-4165(83)90198-8. [DOI] [PubMed] [Google Scholar]
  3. Dettmer R., Kärgel H. J., Stahl J. Preparation of 40S ribosomal subunit proteins of rat liver by combination of ion exchange chromatography with reversed phase liquid chromatography. Biomed Biochim Acta. 1984;43(7):937–945. [PubMed] [Google Scholar]
  4. Ferris R. J., Cowgill C. A., Traut R. R. Separation of ribosomal proteins from Escherichia coli and rabbit reticulocytes using reverse-phase high-performance liquid chromatography. Biochemistry. 1984 Jul 17;23(15):3434–3442. doi: 10.1021/bi00310a009. [DOI] [PubMed] [Google Scholar]
  5. Hardy S. J., Kurland C. G., Voynow P., Mora G. The ribosomal proteins of Escherichia coli. I. Purification of the 30S ribosomal proteins. Biochemistry. 1969 Jul;8(7):2897–2905. doi: 10.1021/bi00835a031. [DOI] [PubMed] [Google Scholar]
  6. Higo K., Otaka E. Isolation and characterization of fourteen ribosomal proteins from small subunits of yeast. Biochemistry. 1979 Sep 18;18(19):4191–4196. doi: 10.1021/bi00586a024. [DOI] [PubMed] [Google Scholar]
  7. Kaltschmidt E., Wittmann H. G. Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Anal Biochem. 1970 Aug;36(2):401–412. doi: 10.1016/0003-2697(70)90376-3. [DOI] [PubMed] [Google Scholar]
  8. Kamp R. M., Bosserhoff A., Kamp D., Wittmann-Liebold B. Application of high-performance liquid chromatographic techniques to the separation of ribosomal proteins of different organisms. J Chromatogr. 1984 Dec 28;317:181–192. doi: 10.1016/s0021-9673(01)91658-9. [DOI] [PubMed] [Google Scholar]
  9. Kamp R. M., Wittmann-Liebold B. Purification of Escherichia coli 50 S ribosomal proteins by high performance liquid chromatography. FEBS Lett. 1984 Feb 13;167(1):59–63. doi: 10.1016/0014-5793(84)80832-7. [DOI] [PubMed] [Google Scholar]
  10. Kamp R. M., Yao Z. J., Bosserhoff A., Wittmann-Liebold B. Purification of Escherichia coli 30S ribosomal proteins by high performance liquid chromatography. Hoppe Seylers Z Physiol Chem. 1983 Dec;364(12):1777–1793. doi: 10.1515/bchm2.1983.364.2.1777. [DOI] [PubMed] [Google Scholar]
  11. Kerlavage A. R., Hasan T., Cooperman B. S. Reverse phase high performance liquid chromatography of Escherichia coli ribosomal proteins: standardization of 70 S, 50 S, and 30 S protein chromatograms. J Biol Chem. 1983 May 25;258(10):6313–6318. [PubMed] [Google Scholar]
  12. Kerlavage A. R., Kahan L., Cooperman B. S. Reverse-phase high-performance liquid chromatography of Escherichia coli ribosomal small subunit proteins. Anal Biochem. 1982 Jul 1;123(2):342–348. doi: 10.1016/0003-2697(82)90456-0. [DOI] [PubMed] [Google Scholar]
  13. Kerlavage A. R., Weitzmann C. J., Hasan T., Cooperman B. S. Reversed-phase high-performance liquid chromatography of Escherichia coli ribosomal proteins. Characteristics of the separation of a complex protein mixture. J Chromatogr. 1983 Aug 26;266:225–237. doi: 10.1016/s0021-9673(01)90896-9. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  16. Otaka E., Higo K., Osawa S. Isolation of seventeen proteins and amino-terminal amino acid sequences of eight proteins from cytoplasmic ribosomes of yeast. Biochemistry. 1982 Sep 14;21(19):4545–4550. doi: 10.1021/bi00262a005. [DOI] [PubMed] [Google Scholar]
  17. Threadgill G. J., Conrad R. C., Changchien L. M., Cannon M., Craven G. R. Application of high-performance liquid chromatography to the purification and characterization of ribosomal protein L3 from trichodermin-resistant yeast mutants. Biochem J. 1986 Jul 15;237(2):421–426. doi: 10.1042/bj2370421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tsurugi K., Collatz E., Todokoro K., Wool I. G. Isolation of eukaryotic ribosomal proteins. Purification and characterization of 60 S ribosomal subunit proteins L3, L6, L7', L8, L10, L15, L17, L18, L19, L23', L25, L27', L28, L29, L31, L32, L34, L35, L36, L36', and L37'. J Biol Chem. 1977 Jun 10;252(11):3961–3969. [PubMed] [Google Scholar]

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