Chaotic advection mixer for capturing transient states of diverse biological macromolecular systems with time-resolved small-angle X-ray scattering

Zielinski K, Katz A, Calvey G, Pabit S, Milano S, Aplin C, San Emeterio J, Cerione R, Pollack L
IUCrJ 10(3):363-375 (2023 Apr 28)

doi: 10.1107/S2052252523003482

Submitted to SASBDB: 2023 Jan 23 Published in SASBDB: 2023 May 4

SASDRE3 – Tissue Transglutaminase + Ca: Time-resolved 0ms Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 4.1 nm Dmax 16.0 nm VolumePorod 150 nm3

SASDRF3 – Tissue Transglutaminase + Ca: Time-resolved 32 ms Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 4.1 nm Dmax 17.0 nm VolumePorod 170 nm3

SASDRG3 – Tissue Transglutaminase + Ca: Time-resolved 63 ms Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 4.1 nm Dmax 18.0 nm VolumePorod 155 nm3

SASDRH3 – Tissue Transglutaminase + Ca: Time-resolved 100 ms Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 4.1 nm Dmax 20.0 nm VolumePorod 190 nm3

SASDRJ3 – Tissue Transglutaminase + Ca: Time-resolved 316 ms Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 4.5 nm Dmax 24.0 nm VolumePorod 200 nm3

SASDRK3 – Tissue Transglutaminase + Ca: Time-resolved 631 ms Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 4.6 nm Dmax 26.0 nm VolumePorod 215 nm3

SASDRL3 – Tissue Transglutaminase + Ca: Time-resolved 1500 ms Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 4.6 nm Dmax 28.0 nm VolumePorod 230 nm3

SASDRM3 – Tissue Transglutaminase + Ca: 5min Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Sep 29 RgGuinier 5.0 nm Dmax 27.0 nm VolumePorod 241 nm3

SASDRN3 – Tissue Transglutaminase + Ca: 10min Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Nov 19 RgGuinier 5.5 nm Dmax 26.0 nm VolumePorod 300 nm3

SASDRP3 – Tissue Transglutaminase + Ca: 30min Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2021 Sep 29 RgGuinier 6.9 nm Dmax 31.5 nm VolumePorod 490 nm3

SASDRQ3 – Trypsin + Aprotinin: Time-Resolved 10 ms Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 1.9 nm Dmax 5.3 nm VolumePorod 23 nm3

SASDRR3 – Trypsin + Aprotinin: Time-Resolved 30 ms Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 1.9 nm Dmax 5.3 nm VolumePorod 28 nm3

SASDRS3 – Trypsin + Aprotinin: Time-Resolved 100 ms Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 1.8 nm Dmax 5.3 nm VolumePorod 30 nm3

SASDRT3 – Trypsin + Aprotinin: Time-Resolved 400 ms Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 1.9 nm Dmax 5.3 nm VolumePorod 27 nm3

SASDRU3 – Trypsin + Aprotinin: Time-Resolved 630 ms Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 1.9 nm Dmax 5.3 nm VolumePorod 25 nm3

SASDRV3 – Trypsin + Aprotinin: Time-Resolved 1000 ms Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 1.8 nm Dmax 5.3 nm VolumePorod 28 nm3

SASDRW3 – Trypsin + Aprotinin: Time-Resolved 2000 ms Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.0 nm Dmax 5.3 nm VolumePorod 29 nm3

SASDRX3 – Trypsin + Aprotinin: Complex equilibrium Sample: Serine protease 1 monomer, 26 kDa Bos taurus protein Pancreatic trypsin inhibitor monomer, 11 kDa Bos taurus protein Buffer: 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH: 7 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 1.9 nm Dmax 5.3 nm VolumePorod 31 nm3

SASDRY3 – Kenics GAC rRNA + Mg: Time-Resolved 10 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.2 nm Dmax 10.0 nm VolumePorod 28 nm3

SASDRZ3 – Kenics GAC rRNA + Mg: Time-Resolved 32 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.2 nm Dmax 10.0 nm VolumePorod 27 nm3

SASDR24 – Kenics GAC rRNA + Mg: Time-Resolved 63 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.2 nm Dmax 9.5 nm VolumePorod 27 nm3

SASDR34 – Kenics GAC rRNA + Mg: Time-Resolved 100 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.2 nm Dmax 10.0 nm VolumePorod 28 nm3

SASDR44 – Kenics GAC rRNA + Mg: Time-Resolved 316 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.2 nm Dmax 12.0 nm VolumePorod 27 nm3

SASDR54 – Kenics GAC rRNA + Mg: Time-Resolved 631 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.3 nm Dmax 9.8 nm VolumePorod 26 nm3

SASDR64 – Kenics GAC rRNA + Mg: Time-Resolved 1000 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.3 nm Dmax 11.0 nm VolumePorod 26 nm3

SASDR74 – Kenics GAC rRNA + Mg: Equilibrium initial Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.4 nm Dmax 10.0 nm VolumePorod 32 nm3

SASDR84 – Kenics GAC rRNA + Mg: Equilibrium final Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.2 nm Dmax 10.0 nm VolumePorod 26 nm3

SASDR94 – GAC rRNA + L11 Protein: Time-Resolved 30 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.4 nm Dmax 12.0 nm VolumePorod 34 nm3

SASDRA4 – GAC rRNA + L11 Protein: Time-Resolved 50 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.5 nm Dmax 13.5 nm VolumePorod 36 nm3

SASDRB4 – GAC rRNA + L11 Protein: Time-Resolved 63 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.4 nm Dmax 11.5 nm VolumePorod 33 nm3

SASDRC4 – GAC rRNA + L11 Protein: Time-Resolved 100 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.4 nm Dmax 11.5 nm VolumePorod 35 nm3

SASDRD4 – GAC rRNA + L11 Protein: Time-Resolved 200 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.5 nm Dmax 12.5 nm VolumePorod 37 nm3

SASDRE4 – GAC rRNA + L11 Protein: Time-Resolved 316 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.5 nm Dmax 10.0 nm VolumePorod 38 nm3

SASDRF4 – GAC rRNA + L11 Protein: Time-Resolved 631 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.5 nm Dmax 13.0 nm VolumePorod 38 nm3

SASDRG4 – GAC rRNA + L11 Protein: Time-Resolved 1000 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.5 nm Dmax 12.5 nm VolumePorod 40 nm3

SASDRH4 – GAC rRNA + L11 Protein: Time-Resolved 2000 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2018 May 9 RgGuinier 2.6 nm Dmax 12.8 nm VolumePorod 39 nm3

SASDRJ4 – GAC rRNA + L11 Protein: SEC-SAXS final Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM Na-MOPSO, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2017 Nov 19 RgGuinier 2.6 nm Dmax 13.0 nm VolumePorod 48 nm3

SASDRK4 – Diffusive GAC rRNA + Mg: Time-Resolved 10 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM sodium cacodylate, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2016 Dec 9 RgGuinier 2.2 nm Dmax 12.0 nm VolumePorod 29 nm3

SASDRL4 – Diffusive GAC rRNA + Mg: Time-Resolved 30 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM sodium cacodylate, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2016 Dec 9 RgGuinier 2.2 nm Dmax 12.0 nm VolumePorod 29 nm3

SASDRM4 – Diffusive GAC rRNA + Mg: Time-Resolved 100 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM sodium cacodylate, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2016 Dec 9 RgGuinier 2.2 nm Dmax 9.0 nm VolumePorod 29 nm3

SASDRN4 – Diffusive GAC rRNA + Mg: Time-Resolved 300 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM sodium cacodylate, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2016 Dec 9 RgGuinier 2.2 nm Dmax 12.0 nm VolumePorod 29 nm3

SASDRP4 – Diffusive GAC rRNA + Mg: Time-Resolved 1000 ms Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM sodium cacodylate, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2016 Dec 9 RgGuinier 2.4 nm Dmax 13.0 nm VolumePorod 30 nm3

SASDRQ4 – Diffusive GAC rRNA + Mg: Equilibrium initial Sample: 58 nucleotide RNA L11-binding domain from E. coli 23S rRNA monomer, 19 kDa Escherichia coli RNA Buffer: 10 mM sodium cacodylate, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2016 Dec 9 RgGuinier 2.4 nm Dmax 10.0 nm VolumePorod 34 nm3

SASDRR4 – Diffusive GAC rRNA + Mg: Equilibrium final Sample: 50S ribosomal protein L11 monomer, 16 kDa Thermus thermophilus protein Buffer: 10 mM sodium cacodylate, 100 mM KCl, pH: 6.5 Experiment: SAXS data collected at G1, Cornell High Energy Synchrotron Source (CHESS) on 2016 Dec 9 RgGuinier 2.2 nm Dmax 11.0 nm VolumePorod 27 nm3