Mapping proteins of the 50S subunit from Escherichia coli ribosomes

• Published in: Biochimica et biophysica acta Vol. 1520; no. 1; pp. 7 - 20
• Main Authors: Willumeit, Regine, Diedrich, Gundo, Forthmann, Stefan, Beckmann, Jörn, May, Roland P, Stuhrmann, Heinrich B, Nierhaus, Knud H
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 30.07.2001
• Subjects: Crystallography; Deuterium; Electrophoresis, Polyacrylamide Gel; Escherichia coli - chemistry; Models, Molecular; Neutron scattering; Neutrons; Protons; Ribosomal protein; Ribosomal Proteins - analysis; Ribosomal Proteins - chemistry; Ribosomes - chemistry; RNA, Ribosomal - isolation & purification; Scattering, Radiation; Spin-contrast variation; EHBA, sodium bis(2-ethyl-2-hydroxybutyrato)oxo-chromate(V) monohydrate {Na[Cr(C 6H 10O 3) 2]·H 2O}; Neutron scattering; IEM, immunoelectron microscopy; DNP, dynamic nuclear polarisation; SANS, small angle neutron scattering; CP, central protuberance; Ribosomal protein; Spin-contrast variation; TP50, total proteins of the 50S ribosomal subunit
• ISSN: 0167-4781; 0006-3002; 1879-2634
• DOI: 10.1016/S0167-4781(01)00245-7

Mapping of protein positions in the ribosomal subunits was first achieved for the 30S subunit by means of neutron scattering about 15 years ago. Since the 50S subunit is almost twice as large as the 30S subunit and consists of more proteins, it was difficult to apply classical contrast variation techniques for the localisation of the proteins. Polarisation dependent neutron scattering (spin-contrast variation) helped to overcome this restriction. Here a map of 14 proteins within the 50S subunit from Escherichia coli ribosomes is presented including the proteins L17 and L20 that are not present in archeal ribosomes. The results are compared with the recent crystallographic map of the 50S subunit from the archea Haloarcula marismortui.