Isolation of Ribosomal Subunits Containing Intact RRNA from Human Placenta: Estimation of Functional Activity of 80S Ribosomes

Overview

Journal Anal Biochem

Publisher Elsevier

Specialty Biochemistry

Date 1991 Nov 1

PMID 1799204

Citations 27

Authors

N B Matasova

S V Myltseva

M A Zenkova

D M Graifer

S N Vladimirov

G G Karpova

Affiliations

Soon will be listed here.

Abstract

We have elaborated a method for the isolation of ribosomal subunits from fresh unfrozen human placenta containing intact rRNA and a complete set of ribosomal proteins. Activity of 80S ribosomes obtained by reassociation of 40S and 60S subunits in nonenzymatic poly(U)-dependent binding of Phe-tRNA(Phe) was equal to 80% (above 1.5 mol [14C]Phe-tRNA(Phe) is coupled to 1 mol of ribosomes). The activity of 80S ribosomes in poly(U)-directed synthesis of polyphenylalanine was tested in a polysome-free protein-synthesizing system from rabbit reticulocytes. About 100 mol of phenylalanine residue was polymerized by a mole of ribosomes at a rate of 0.83 residues per minute in this system (2 h, 37 degrees C).

Citing Articles

The flexible N-terminal motif of uL11 unique to eukaryotic ribosomes interacts with P-complex and facilitates protein translation.

Yang L, Lee K, Yu C, Imai H, Choi A, Banfield D Nucleic Acids Res. 2022; 50(9):5335-5348.

PMID: 35544198 PMC: 9122527. DOI: 10.1093/nar/gkac292.

Lyophyllin, a Mushroom Protein from the Peptidase M35 Superfamily Is an RNA N-Glycosidase.

Lu J, Shi W, Xiao M, Tang Y, Zheng Y, Shaw P Int J Mol Sci. 2021; 22(21).

PMID: 34769028 PMC: 8584072. DOI: 10.3390/ijms222111598.

Two alternative conformations of mRNA in the human ribosome during elongation and termination of translation as revealed by EPR spectroscopy.

Bulygin K, Timofeev I, Malygin A, Graifer D, Meschaninova M, Venyaminova A Comput Struct Biotechnol J. 2021; 19:4702-4710.

PMID: 34504663 PMC: 8390954. DOI: 10.1016/j.csbj.2021.08.024.

mRNA regions where 80S ribosomes pause during translation elongation in vivo interact with protein uS19, a component of the decoding site.

Babaylova E, Gopanenko A, Bulygin K, Tupikin A, Kabilov M, Malygin A Nucleic Acids Res. 2019; 48(2):912-923.

PMID: 31802126 PMC: 6954443. DOI: 10.1093/nar/gkz1145.

Exploring the interactions of short RNAs with the human 40S ribosomal subunit near the mRNA entry site by EPR spectroscopy.

Malygin A, Krumkacheva O, Graifer D, Timofeev I, Ochkasova A, Meschaninova M Nucleic Acids Res. 2019; 47(22):11850-11860.

PMID: 31724718 PMC: 7145563. DOI: 10.1093/nar/gkz1039.