Distinct conformational states enable transglutaminase 2 to promote cancer cell survival versus cell death.

Aplin C, Zielinski KA, Pabit S, Ogunribido D, Katt WP, Pollack L, Cerione RA, Milano SK
Commun Biol 7(1):982 (2024 Aug 13)

PMID: 39134806 doi: 10.1038/s42003-024-06672-x

Submitted to SASBDB: 2023 Nov 2 Published in SASBDB: 2024 Jul 24

SASDTL3 – Tissue Transglutaminase: 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 2022 Nov 17 RgGuinier 4.2 nm Dmax 20.0 nm VolumePorod 148 nm3

SASDTM3 – Tissue Transglutaminase + 25 µM GTP 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 2022 Nov 17 RgGuinier 3.8 nm Dmax 17.5 nm VolumePorod 132 nm3

SASDTN3 – Tissue Transglutaminase + 50 µM GTP 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 2022 Nov 17 RgGuinier 3.7 nm Dmax 17.5 nm VolumePorod 124 nm3

SASDTP3 – Tissue Transglutaminase + 100 µM GTP 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 2022 Nov 17 RgGuinier 3.6 nm Dmax 18.0 nm VolumePorod 121 nm3

SASDTQ3 – Tissue Transglutaminase + 250 µM GTP 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 2022 Nov 17 RgGuinier 3.5 nm Dmax 17.5 nm VolumePorod 120 nm3

SASDTR3 – Tissue Transglutaminase + 500 µM GTP 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 2022 Nov 17 RgGuinier 3.4 nm Dmax 18.0 nm VolumePorod 118 nm3

SASDTS3 – Tissue Transglutaminase + 1 mM GTP 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 2022 Nov 17 RgGuinier 3.4 nm Dmax 17.0 nm VolumePorod 115 nm3

SASDTT3 – Tissue Transglutaminase + 2 mM GTP 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 2022 Nov 17 RgGuinier 3.3 nm Dmax 17.0 nm VolumePorod 111 nm3

SASDTU3 – Tissue Transglutaminase + 5 mM GTP: Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, 5 mM GTP, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2022 Nov 17 RgGuinier 3.2 nm Dmax 14.0 nm VolumePorod 108 nm3

SASDTV3 – Tissue Transglutaminase + 5 mM GDP 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 2022 Nov 17 RgGuinier 3.5 nm Dmax 17.5 nm VolumePorod 120 nm3

SASDTW3 – Tissue Transglutaminase + 250 µM CaCl2 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 2022 Nov 17 RgGuinier 4.3 nm Dmax 21.0 nm VolumePorod 160 nm3

SASDTX3 – Tissue Transglutaminase + 500 µM CaCl2 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 2022 Nov 17 RgGuinier 4.3 nm Dmax 20.0 nm VolumePorod 158 nm3

SASDTY3 – Tissue Transglutaminase + 1 mM CaCl2 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 2022 Nov 17 RgGuinier 4.5 nm Dmax 24.0 nm VolumePorod 180 nm3

SASDTZ3 – Tissue Transglutaminase + 2 mM CaCl2 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 2022 Nov 17 RgGuinier 5.3 nm Dmax 25.5 nm VolumePorod 234 nm3

SASDT24 – Tissue Transglutaminase + 50 µM inhibitor LM11 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 2022 Nov 17 RgGuinier 4.3 nm Dmax 20.0 nm VolumePorod 158 nm3

SASDT34 – Tissue Transglutaminase + 50 µM inhibitor LM11 + 250 µM CaCl2 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 2022 Nov 17 RgGuinier 4.2 nm Dmax 19.0 nm VolumePorod 155 nm3

SASDT44 – Tissue Transglutaminase + 50 µM inhibitor LM11 + 500 µM CaCl2 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 2022 Nov 17 RgGuinier 4.9 nm Dmax 25.0 nm VolumePorod 191 nm3

SASDT54 – Tissue Transglutaminase + 50 µM inhibitor LM11 + 1 mM CaCl2 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 2022 Nov 17 RgGuinier 4.7 nm Dmax 28.0 nm VolumePorod 198 nm3

SASDT64 – Tissue Transglutaminase + 50 µM inhibitor LM11 + 2 mM CaCl2 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 2022 Nov 17 RgGuinier 5.2 nm Dmax 25.0 nm VolumePorod 236 nm3

SASDT74 – Tissue Transglutaminase + CaCl2 + inhibitor LM11: Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, 2 mM CaCl2, 50 µM LM11, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2022 Nov 17 RgGuinier 6.0 nm Dmax 36.0 nm VolumePorod 431 nm3

SASDT84 – Tissue Transglutaminase + inhibitor LM11 + CaCl2: 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 2022 Nov 17 RgGuinier 4.6 nm Dmax 32.0 nm VolumePorod 242 nm3

SASDT94 – Tissue Transglutaminase + inhibitor LM11 + CaCl2: 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 2022 Nov 17 RgGuinier 4.8 nm Dmax 25.0 nm VolumePorod 203 nm3

SASDTA4 – Tissue Transglutaminase + inhibitor LM11 + CaCl2: 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 2022 Nov 17 RgGuinier 5.7 nm Dmax 33.0 nm VolumePorod 305 nm3

SASDTB4 – Tissue Transglutaminase + CaCl2 + inhibitor LM11 + GTP: Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, 2 mM CaCl2, 50 µM LM11, 5 mM GTP, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2022 Nov 17 RgGuinier 6.2 nm Dmax 33.0 nm VolumePorod 277 nm3

SASDTC4 – Tissue Transglutaminase + 50 µM inhibitor 5826 (1% DMSO) 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 2022 Nov 17 RgGuinier 4.1 nm Dmax 19.0 nm VolumePorod 144 nm3

SASDTD4 – Tissue Transglutaminase + 50 µM inhibitor 5826 (2% DMSO) 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 2022 Nov 17 RgGuinier 3.9 nm Dmax 18.5 nm VolumePorod 134 nm3

SASDTE4 – Tissue Transglutaminase + 100 µM inhibitor 5826 (2% DMSO) 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 2022 Nov 17 RgGuinier 3.8 nm Dmax 17.6 nm VolumePorod 130 nm3

SASDTF4 – Tissue Transglutaminase + CaCl2 + inhibitor 5826: Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, 2 mM CaCl2, 50 µM 5826 inhibitor, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2022 Nov 17 RgGuinier 6.7 nm Dmax 39.0 nm VolumePorod 450 nm3

SASDTG4 – Tissue Transglutaminase + CaCl2 + inhibitor 5826 + GTP: Equilibrium Sample: Protein-glutamine gamma-glutamyltransferase 2, 77 kDa Homo sapiens protein Buffer: 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, 2 mM CaCl2, 50 µM 5826, 5 mM GTP, pH: 7.5 Experiment: SAXS data collected at ID7A1 BioSAXS / HP-Bio Beamline, Cornell High Energy Synchrotron Source (CHESS) on 2022 Nov 17 RgGuinier 6.0 nm Dmax 29.0 nm VolumePorod 248 nm3

SASDTH4 – Tissue Transglutaminase + 1% DMSO 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 2022 Nov 17 RgGuinier 4.0 nm Dmax 18.5 nm VolumePorod 139 nm3

SASDTJ4 – Tissue Transglutaminase + 2% DMSO 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 2022 Nov 17 RgGuinier 3.9 nm Dmax 18.5 nm VolumePorod 133 nm3

SASDTK4 – Tissue Transglutaminase + 5% DMSO 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 2022 Nov 17 RgGuinier 3.9 nm Dmax 18.5 nm VolumePorod 133 nm3

SASDTL4 – Tissue Transglutaminase + 10% DMSO 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 2022 Nov 17 RgGuinier 4.1 nm Dmax 21.5 nm VolumePorod 148 nm3

SASDTM4 – Tissue Transglutaminase R580K mutant Sample: Protein-glutamine gamma-glutamyltransferase 2 (R580K), 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 2022 Nov 17 RgGuinier 6.1 nm Dmax 32.0 nm VolumePorod 488 nm3

SASDTN4 – Tissue Transglutaminase R580K mutant + 100 µM GTP Sample: Protein-glutamine gamma-glutamyltransferase 2 (R580K), 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 2022 Nov 17 RgGuinier 5.9 nm Dmax 27.0 nm VolumePorod 381 nm3

SASDTP4 – Tissue Transglutaminase R580K mutant + 1 mM GTP Sample: Protein-glutamine gamma-glutamyltransferase 2 (R580K), 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 2022 Nov 17 RgGuinier 5.7 nm Dmax 30.0 nm VolumePorod 283 nm3

SASDTQ4 – Tissue Transglutaminase R580K mutant + 2 mM GTP Sample: Protein-glutamine gamma-glutamyltransferase 2 (R580K), 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 2022 Nov 17 RgGuinier 5.3 nm Dmax 25.0 nm VolumePorod 264 nm3

SASDTR4 – Tissue Transglutaminase R580K mutant + 5 mM GTP Sample: Protein-glutamine gamma-glutamyltransferase 2 (R580K), 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 2022 Nov 17 RgGuinier 5.2 nm Dmax 25.0 nm VolumePorod 243 nm3