Domain crossover in the reductase subunit of NADPH-dependent assimilatory sulfite reductase.

Walia N, Murray DT, Garg Y, He H, Weiss KL, Nagy G, Elizabeth Stroupe M
J Struct Biol 215(4):108028 (2023 Sep 12)

PMID: 37704014 doi: 10.1016/j.jsb.2023.108028

Submitted to SASBDB: 2023 Apr 30 Published in SASBDB: 2023 Sep 24

SASDS22 – Sulfite reductase flavoprotein crosslinked octamer Sample: Sulfite reductase [NADPH] flavoprotein alpha-component octamer, 530 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jun 24 RgGuinier 8.4 nm Dmax 23.0 nm VolumePorod 767 nm3

SASDS32 – Reconstituted sulfite reductase crosslinked dodecamer with partially deuterated hemoprotein in 86% D2O Sample: Sulfite reductase [NADPH] hemoprotein beta-component tetramer, 256 kDa Escherichia coli (strain … protein Sulfite reductase [NADPH] flavoprotein alpha-component (E121C, C162T, C552S, N556C) octamer, 566 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2023 Apr 24 RgGuinier 7.5 nm Dmax 22.6 nm VolumePorod 823 nm3

SASDS42 – Reconstituted sulfite reductase crosslinked dodecamer with partially deuterated hemoprotein in 41% D2O Sample: Sulfite reductase [NADPH] hemoprotein beta-component tetramer, 256 kDa Escherichia coli (strain … protein Sulfite reductase [NADPH] flavoprotein alpha-component (E121C, C162T, C552S, N556C) octamer, 566 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Aug 24 RgGuinier 11.1 nm Dmax 32.4 nm VolumePorod 138 nm3

SASDS52 – Sulfite reductase crosslinked dodecamer Sample: Sulfite reductase [NADPH] hemoprotein beta-component tetramer, 256 kDa Escherichia coli (strain … protein Sulfite reductase [NADPH] flavoprotein alpha-component (E121C, C162T, C552S, N556C) octamer, 566 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jul 15 RgGuinier 10.3 nm Dmax 31.0 nm VolumePorod 1170 nm3

SASDS62 – Sulfite reductase flavoprotein truncated linker (ΔAAPSQS) octamer Sample: Sulfite reductase [NADPH] flavoprotein alpha-component (Δ212-217) octamer, 526 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jun 25 RgGuinier 13.7 nm Dmax 25.5 nm VolumePorod 961 nm3

SASDS72 – Sulfite reductase truncated linker (ΔAAPSQS) dodecamer Sample: Sulfite reductase [NADPH] hemoprotein beta-component tetramer, 256 kDa Escherichia coli (strain … protein Sulfite reductase [NADPH] flavoprotein alpha-component (Δ212-217) octamer, 528 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jun 26 RgGuinier 10.8 nm Dmax 30.3 nm VolumePorod 1260 nm3

SASDS82 – Reconstituted sulfite reductase crosslinked dodecamer with partially deuterated hemoprotein in 0% D2O Sample: Sulfite reductase [NADPH] flavoprotein alpha-component (E121C, C162T, C552S, N556C) octamer, 566 kDa Escherichia coli (strain … protein Sulfite reductase [NADPH] hemoprotein beta-component tetramer, 256 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jul 2 RgGuinier 9.5 nm Dmax 28.3 nm VolumePorod 1010 nm3

SASDS92 – Reconstituted sulfite reductase crosslinked dodecamer with partially deuterated hemoprotein in 100% D2O Sample: Sulfite reductase [NADPH] hemoprotein beta-component tetramer, 256 kDa Escherichia coli (strain … protein Sulfite reductase [NADPH] flavoprotein alpha-component (E121C, C162T, C552S, N556C) octamer, 566 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jun 27 RgGuinier 8.2 nm Dmax 25.1 nm VolumePorod 839 nm3

SASDSA2 – Sulfite reductase flavoprotein reduced with 10 molar equivalents sodium dithionite Sample: Sulfite reductase [NADPH] flavoprotein alpha-component octamer, 530 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jul 15 RgGuinier 7.9 nm Dmax 27.6 nm VolumePorod 1060 nm3

SASDSB2 – Sulfite reductase flavoprotein crosslinked reduced with 10 molar equivalents sodium dithionite Sample: Sulfite reductase [NADPH] flavoprotein alpha-component (E121C, C162T, C552S, N556C) octamer, 566 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jul 15 RgGuinier 8.0 nm Dmax 25.3 nm VolumePorod 827 nm3

SASDSC2 – Sulfite reductase truncated linker (ΔAAPSQS) flavoprotein reduced with 10 molar equivalents sodium dithionite Sample: Sulfite reductase [NADPH] flavoprotein alpha-component (Δ212-217) octamer, 526 kDa Escherichia coli (strain … protein Buffer: 50 mM potassium phosphate, 100 mM NaCl, 1 mM EDTA, pH: 7.8 Experiment: SANS data collected at EQ-SANS, Spallation Neutron Source on 2022 Jul 15 RgGuinier 7.6 nm Dmax 25.0 nm VolumePorod 952 nm3