Frozen spin targets in ribosomal structure research

Solution scattering structural analysis of the 70 S Escherichia coli ribosome by contrast variation. I. invariants and validation of electron microscopy models 1 1Edited by M. F. Moody

“…Polarized neutron scattering measurements from nuclear spin polarized samples (spin-dependent contrast variation) were performed at the SANS-1 instrument of the FRG-1 reactor at the GKSS Research Center, Geesthacht (Stuhrmann, 1991;Stuhrmann et al, 1995). The samples were frozen solutions of selectively deuterated 70 S ribosomes (Table 1; ribosome concentration 7 to 11 mg/ml) in a mixture of perdeuterated glycerol and heavy water (11/9) containing 100 mM imidazole (pH 7.6; 0 C), 10 mM MgCl 2 , 100 mM KCl and 20 mM sodium bis(2-ethyl-2-hydroxybutyrato)-oxochromate(V)-monohydrate.…”

Solution scattering structural analysis of the 70 S Escherichia coli ribosome by contrast variation. I. invariants and validation of electron microscopy models 1 1Edited by M. F. Moody

“…Polarized neutron scattering measurements from nuclear spin polarized samples (spin-dependent contrast variation) were performed at the SANS-1 instrument of the FRG-1 reactor at the GKSS Research Center, Geesthacht (Stuhrmann, 1991;Stuhrmann et al, 1995). The samples were frozen solutions of selectively deuterated 70 S ribosomes (Table 1; ribosome concentration 7 to 11 mg/ml) in a mixture of perdeuterated glycerol and heavy water (11/9) containing 100 mM imidazole (pH 7.6; 0 C), 10 mM MgCl 2 , 100 mM KCl and 20 mM sodium bis(2-ethyl-2-hydroxybutyrato)-oxochromate(V)-monohydrate.…”

Direct localization of the tRNAs within the elongating ribosome by means of neutron scattering (proton-spin contrast-variation)

“…Spectra were taken for the unpolarized target and with either parallel or antiparallel aligned proton spins relative to the spin of the incoming neutrons. The collected data were processed according to standard corrections (May et al, 1992) and deconvoluted into the three basic scattering functions [UU], [UH] and [HH] for the pre and the post-translocational complexes (Ibel & Stuhrmann, 1975;Stuhrmann, 1991;Figure 2 (Ibel & Stuhrmann, 1975) PRE, pre-translocational complex; POST, post-translocational complex. The distance between the centre of mass of the protonated (tRNA) 2 mRNA complex and that of the ribosome could be determined without any knowledge of the ribosomal structure. However, information on the position of the mass centre of the protonated complex within the deuterated ribosome is most readily extracted from the data by referring to a model of the 70 S ribosome.…”

A Map of Protein-rRNA Distribution in the 70 SEscherichia coli Ribosome

“…The 70 S samples of the E. coli ribosome were made by in vitro association of selectively deuterated ribosomal subunits and checked for structural integrity and biological activity. The neutron experiments were performed at the Risø Laboratory (Roskilde, Denmark (20)) and at the GKSS Research Center (Geesthacht, Germany (21,22)). Using two sample-detector distances ("low angle" and "high angle" setting), the range of momentum transfer 0.09 Ͻ s Ͻ 2.3 nm Ϫ1 was covered (s ϭ (4/)sin, where is the wavelength, and 2 is the scattering angle).…”

Solution scattering structural analysis of the 70 S Escherichia coli ribosome by contrast variation. I. invariants and validation of electron microscopy models 1 1Edited by M. F. Moody

“…Polarized neutron scattering measurements from nuclear spin polarized samples (spin-dependent contrast variation) were performed at the SANS-1 instrument of the FRG-1 reactor at the GKSS Research Center, Geesthacht (Stuhrmann, 1991;Stuhrmann et al, 1995). The samples were frozen solutions of selectively deuterated 70 S ribosomes (Table 1; ribosome concentration 7 to 11 mg/ml) in a mixture of perdeuterated glycerol and heavy water (11/9) containing 100 mM imidazole (pH 7.6; 0 C), 10 mM MgCl 2 , 100 mM KCl and 20 mM sodium bis(2-ethyl-2-hydroxybutyrato)-oxochromate(V)-monohydrate.…”

Direct localization of the tRNAs within the elongating ribosome by means of neutron scattering (proton-spin contrast-variation)

“…Spectra were taken for the unpolarized target and with either parallel or antiparallel aligned proton spins relative to the spin of the incoming neutrons. The collected data were processed according to standard corrections (May et al, 1992) and deconvoluted into the three basic scattering functions [UU], [UH] and [HH] for the pre and the post-translocational complexes (Ibel & Stuhrmann, 1975;Stuhrmann, 1991;Figure 2 (Ibel & Stuhrmann, 1975) PRE, pre-translocational complex; POST, post-translocational complex. The distance between the centre of mass of the protonated (tRNA) 2 mRNA complex and that of the ribosome could be determined without any knowledge of the ribosomal structure. However, information on the position of the mass centre of the protonated complex within the deuterated ribosome is most readily extracted from the data by referring to a model of the 70 S ribosome.…”

A Map of Protein-rRNA Distribution in the 70 SEscherichia coli Ribosome

“…The 70 S samples of the E. coli ribosome were made by in vitro association of selectively deuterated ribosomal subunits and checked for structural integrity and biological activity. The neutron experiments were performed at the Risø Laboratory (Roskilde, Denmark (20)) and at the GKSS Research Center (Geesthacht, Germany (21,22)). Using two sample-detector distances ("low angle" and "high angle" setting), the range of momentum transfer 0.09 Ͻ s Ͻ 2.3 nm Ϫ1 was covered (s ϭ (4/)sin, where is the wavelength, and 2 is the scattering angle).…”

Solution scattering structural analysis of the 70 S Escherichia coli ribosome by contrast variation. I. invariants and validation of electron microscopy models 1 1Edited by M. F. Moody

“…Polarized neutron scattering measurements from nuclear spin polarized samples (spin-dependent contrast variation) were performed at the SANS-1 instrument of the FRG-1 reactor at the GKSS Research Center, Geesthacht (Stuhrmann, 1991;Stuhrmann et al, 1995). The samples were frozen solutions of selectively deuterated 70 S ribosomes (Table 1; ribosome concentration 7 to 11 mg/ml) in a mixture of perdeuterated glycerol and heavy water (11/9) containing 100 mM imidazole (pH 7.6; 0 C), 10 mM MgCl 2 , 100 mM KCl and 20 mM sodium bis(2-ethyl-2-hydroxybutyrato)-oxochromate(V)-monohydrate.…”

Direct localization of the tRNAs within the elongating ribosome by means of neutron scattering (proton-spin contrast-variation)

“…Spectra were taken for the unpolarized target and with either parallel or antiparallel aligned proton spins relative to the spin of the incoming neutrons. The collected data were processed according to standard corrections (May et al, 1992) and deconvoluted into the three basic scattering functions [UU], [UH] and [HH] for the pre and the post-translocational complexes (Ibel & Stuhrmann, 1975;Stuhrmann, 1991;Figure 2 (Ibel & Stuhrmann, 1975) PRE, pre-translocational complex; POST, post-translocational complex. The distance between the centre of mass of the protonated (tRNA) 2 mRNA complex and that of the ribosome could be determined without any knowledge of the ribosomal structure. However, information on the position of the mass centre of the protonated complex within the deuterated ribosome is most readily extracted from the data by referring to a model of the 70 S ribosome.…”

A Map of Protein-rRNA Distribution in the 70 SEscherichia coli Ribosome

“…The 70 S samples of the E. coli ribosome were made by in vitro association of selectively deuterated ribosomal subunits and checked for structural integrity and biological activity. The neutron experiments were performed at the Risø Laboratory (Roskilde, Denmark (20)) and at the GKSS Research Center (Geesthacht, Germany (21,22)). Using two sample-detector distances ("low angle" and "high angle" setting), the range of momentum transfer 0.09 Ͻ s Ͻ 2.3 nm Ϫ1 was covered (s ϭ (4/)sin, where is the wavelength, and 2 is the scattering angle).…”