Home LiteratureArticle Details
PMID: 3475694 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Topography and stoichiometry of acidic proteins in large ribosomal subunits from Artemia salina as determined by crosslinking.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 84 ·No. 16 ·1987-08-00 ·Pages 5580-4

Uchiumi T, Wahba AJ, Traut RR

Abstract

The 60S subunits isolated from Artemia salina ribosomes were treated with the crosslinking reagent 2-iminothiolane under mild conditions. Proteins were extracted and fractions containing crosslinked acidic proteins were obtained by stepwise elution from CM-cellulose. Each fraction was analyzed by "diagonal" (two-dimensional nonreducing-reducing) NaDodSO4/polyacrylamide gel electrophoresis. Crosslinked proteins below the diagonal were radioiodinated and identified by two-dimensional acidic urea-NaDodSO4 gel electrophoresis. Each of the acidic proteins P1 and P2 was crosslinked individually to the same third protein, P0. The fractions containing acidic proteins were also analyzed by two-dimensional nonequilibrium isoelectric focusing-NaDodSO4/polyacrylamide gel electrophoresis. Two crosslinked complexes were observed that coincide in isoelectric positions with monomeric P1 and P2, respectively. Both P1 and P2 appear to form crosslinked homodimers. These results suggest the presence in the 60S subunit of (P1)2 and (P2)2 dimers, each of which is anchored to P0. Protein P0 appears to play the same role as L10 in Escherichia coli ribosomes and may form a pentameric complex with the two dimers in the 60S subunits.

MeSH Terms
Animals Artemia/analysis Cross-Linking Reagents/pharmacology Electrophoresis, Polyacrylamide Gel Imidoesters/pharmacology Isoelectric Focusing Protein Conformation Ribosomal Proteins/analysis
Chemicals
Cross-Linking Reagents Imidoesters Ribosomal Proteins methyl 4-mercaptobutyrimidate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Uchiumi T
Wahba A J
Traut R R
References (37)
37 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Isolation and characterization of the acidic phosphoproteins of 60-S ribosomes from Artemia salina and rat liver.
    Eur J Biochem. 1978 Nov 15;91(2):553-65 PMID: 32034
  3. 50-S ribosomal proteins. Purification and partial characterization of two acidic proteins, A 1 and A 2, isolated from 50-S ribosomes of Escherichia coli.
    Eur J Biochem. 1972 Jan 31;25(1):5-12 PMID: 4554105
  4. Diagonal polyacrylamide-dodecyl sulfate gel electrophoresis for the identification of ribosomal proteins crosslinked with methyl-4-mercaptobutyrimidate.
    Proc Natl Acad Sci U S A. 1974 Oct;71(10):3946-50 PMID: 4610565
  5. Shape properties of proteins L7 and L12 from E. coli ribosomes.
    Biochem Biophys Res Commun. 1974 Nov 6;61(1):178-84 PMID: 4613351
  6. Copies of proteins L7 and L12 and heterogeneity of the large subunit of Escherichia coli ribosome.
    J Mol Biol. 1975 Jun 15;95(1):1-8 PMID: 1097708
  7. The stoichiometry of the ribosomal proteins of Escherichia coli.
    Mol Gen Genet. 1975 Oct 3;140(3):253-74 PMID: 1107798
  8. The ribosomal protein L8 is a complex L7/L12 and L10.
    FEBS Lett. 1976 Apr 15;64(1):135-8 PMID: 773698
  9. Isolation of eukaryotic ribosomal proteins. Purification and characterization of the 60 S ribosomal subunit proteins La, Lb, Lf, P1, P2, L13', L14, L18', L20, and L38.
    J Biol Chem. 1978 Feb 10;253(3):946-55 PMID: 621213
  10. Nucleotide sequence of the ribosomal protein gene cluster adjacent to the gene for RNA polymerase subunit beta in Escherichia coli.
    Proc Natl Acad Sci U S A. 1979 Apr;76(4):1697-701 PMID: 377281
  11. The use of Artemia salina for developmental studies: preparation of embryos, tRNA, ribosomes and initiation factor 2.
    Methods Enzymol. 1979;60:298-311 PMID: 459905
  12. Identification of neighboring protein pairs in rat liver 60S ribosomal subunits cross-linked with dimethyl suberimidate or dimethyl 3,3'-dithiobispropionimidate.
    J Biochem. 1980 Oct;88(4):1033-44 PMID: 7451403
  13. The number of copies of ribosome-bound proteins L7 and L12 required for protein synthesis activity.
    J Biol Chem. 1981 Jan 10;256(1):41-8 PMID: 6108955
  14. Functional role of acidic ribosomal proteins. Interchangeability of proteins from bacterial and eukaryotic cells.
    Biochemistry. 1981 May 26;20(11):3263-6 PMID: 6113843
  15. Identification of proteins at the binding site for protein S1 in 70 S ribosomes and 30 S subunits by cross-linking with 2-iminothiolane.
    J Biol Chem. 1981 Aug 10;256(15):8222-7 PMID: 7021546
  16. Protein topography of the 40 S ribosomal subunit from rabbit reticulocytes shown by cross-linking with 2-iminothiolane.
    J Biol Chem. 1981 Oct 10;256(19):10129-36 PMID: 7275971
  17. The activity of the acidic phosphoproteins from the 80 S rat liver ribosome.
    J Biol Chem. 1982 May 25;257(10):5359-66 PMID: 6121796
  18. The primary structure of the acidic phosphoprotein P2 from rat liver 60 S ribosomal subunits. Comparison with ribosomal 'A' proteins from other species.
    J Biol Chem. 1982 Aug 10;257(15):9189-97 PMID: 7096359
  19. Monoclonal antibodies against eucaryotic ribosomes. Use to characterize a ribosomal protein not previously identified and antigenically related to the acidic phosphoproteins P1/P2.
    J Biol Chem. 1982 Nov 10;257(21):12709-15 PMID: 6182142
  20. Ribosomal proteins L7/L12 of Escherichia coli. Localization and possible molecular mechanism in translation.
    J Mol Biol. 1983 Feb 5;163(4):553-73 PMID: 6133002
  21. Acidic proteins of the large ribosomal subunit in Saccharomyces cerevisiae. Effect of phosphorylation.
    Biochemistry. 1984 Jan 17;23(2):390-6 PMID: 6421316
  22. Cross-linking study on protein topography of rat liver 60 S ribosomal subunits with 2-iminothiolane.
    J Biol Chem. 1985 May 10;260(9):5675-82 PMID: 3988768
  23. Preparation and characterization of two monoclonal antibodies against different epitopes in Escherichia coli ribosomal protein L7/L12.
    J Biol Chem. 1985 Jun 10;260(11):6522-7 PMID: 2581957
  24. Lupus autoantibodies target ribosomal P proteins.
    J Exp Med. 1985 Aug 1;162(2):459-71 PMID: 2410526
  25. Identification of ribosomal protein autoantigens.
    J Immunol. 1985 Oct;135(4):2378-84 PMID: 3897374
  26. Cross-linking of elongation factor 2 to rat-liver ribosomal proteins by 2-iminothiolane.
    Eur J Biochem. 1986 Apr 1;156(1):37-48 PMID: 3956508
  27. Localization of two epitopes of protein L7/L12 to both the body and stalk of the large ribosomal subunit. Immune electron microscopy using monoclonal antibodies.
    J Biol Chem. 1986 May 25;261(15):6924-32 PMID: 2422173
  28. Cross-linking study on localization of the binding site for elongation factor 1 alpha on rat liver ribosomes.
    J Biol Chem. 1986 Jul 25;261(21):9668-71 PMID: 3733692
  29. Identification and chemical synthesis of a ribosomal protein antigenic determinant in systemic lupus erythematosus.
    Proc Natl Acad Sci U S A. 1986 Oct;83(19):7419-23 PMID: 2429305
  30. Protein topography of the 40 S ribosomal subunit from Saccharomyces cerevisiae as shown by chemical cross-linking.
    J Biol Chem. 1986 Oct 25;261(30):14148-53 PMID: 3533916
  31. Small-angle X-ray scattering and crosslinking study of the proteins L7/L12 from Escherichia coli ribosomes.
    FEBS Lett. 1976 Jul 1;66(1):48-51 PMID: 776697
  32. Structure and phosphorylation of an acidic protein from 60S ribosomes and its involvement in elongation factor-2 dependent GTP hydrolysis.
    Biochem Biophys Res Commun. 1977 Aug 8;77(3):989-98 PMID: 197953
  33. High resolution two-dimensional electrophoresis of basic as well as acidic proteins.
    Cell. 1977 Dec;12(4):1133-41 PMID: 23215
  34. A comparison of the alanine-rich sequences of the L7/L12-ribosomal proteins from rat liver, Artemia salina and Escherichia coli, with the amino-terminal region of the alkali light chain A1 from rabbit myosin.
    FEBS Lett. 1978 Feb 15;86(2):282-4 PMID: 342275
  35. Stoichiometry and properties of the complex between ribosomal proteins L7 and L10 in solution.
    FEBS Lett. 1978 Sep 15;93(2):215-8 PMID: 361431
  36. Disulfide cross-linking of Escherichia coli ribosomal proteins with 2-iminothiolane (methyl 4-mercaptobutyrimidate): evidence that the cross-linked protein pairs are formed in the intact ribosomal subunit.
    Biochemistry. 1978 Dec 12;17(25):5406-16 PMID: 365230
  37. Requirement of an Escherichia coli 50 S ribosomal protein component for effective interaction of the ribosome with T and G factors and with guanosine triphosphate.
    J Biol Chem. 1972 Feb 10;247(3):805-14 PMID: 4333515
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.84.16.5580
Published
1987-08-00
Pages
5580-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC298906
Subset
IM
Grants
NIGMS NIH HHS · GM35375 · United States
External Links
PubMed source DOI: 10.1073/pnas.84.16.5580 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2