AR3436A report

1. Sim CN NOESY, aromatic NOESY, CCH-TOCSY were reprocessed and analyzed to evaluate the assignment and structure. Oliver's assignment corrections are confirmed (~100), and additional assignment corrections are found as well (~20). The new assignment has > 120 shifts with (H>0.045ppm, C, N>0.45ppm) compared to the deposited shifts. New assignment and submitted PDB NOESY peak lists (or submitted casd-NMR peak lists) still gave opened structure as deposited one. Sim CN NOESY and aromatic NOESY were re-picked and the new NOESY peak lists and new assignment gave closed structure as CS-Rosetta structure, by CYANA or ASDP. The new peak lists with previous deposited assignment also gave closed structures, by CYANA or ASDP The opened deposited structure became closed after consRosetta refinement, with old constraints and defined loop region. AR3436A report

2. Newcy vs. Newasdp vs. Deposited The structure is closedwith new peak lists and chemical shift

3. Newpeakonly vs. Newshiftonly vs. Deposit The cyana structure is closedwith new peak lists and old chemical shift The cyana structure is openwith old peak lists and new chemical shift

4. Newpeakonlyasdp vs. Newasdp vs. Deposit The asdp structure is closedwith new peak lists and old chemical shift

5. Newcy vs. consRos_dep vs. Deposit The opened deposit structure is closedafter refinement with constraint Rosetta

6. Different Loop Region Definition affect consRos Refinement A, Deposited; B, without loop;C: loop 1-14, 25-27, 29-30, 35-36, 50-54, 73-72, 83-87, 95-97, defined as no-ordered residues by psvs;D: loop 1-14, 22-28, 51-56, 63-67, 85-97,F: New cns structure

7. AR3436A has different sets of peak lists available From Rajee CasdNMR: 15N: 577peaks ali: 1402 peaks, aro: 97 peaks HarvestDB: 15N: 581peaks ali: 1399 peaks, aro: 98 peaks (similar to casd NMR) July2009: 15N: 788peaks ali: 1796 peaks, aro: 166 peaks; find in “mani_home/nmrdata/AR3436A/AR3436A_structure”, not available to public? New peak lists: 15N: 1041 peaks ali: 3451 peaks, aro: 328 peaks Calculation done for July 23th 2009 peaks, old shift Casd peaks, new shifts July 23th 2009 peaks, new shifts July 23th 2009 peaks, new shifts AR3436A Report updated

8. New assignment and submitted PDB NOESY peak lists (or submitted casd-NMR peak lists) still gave relative opened structure , but the hole is smaller than that of the deposited structure. New assignment and old July 23th 2009 peak lists (or submitted casd-NMR peak lists) still gave relative opened structure , but the hole is smaller than those of the deposited structure and 1. Old assignment and old July 23th 2009 peak lists (or submitted casd-NMR peak lists) still gave relative opened structure , but the hole is smaller than those of the deposited structure, 1 and 2. Sim CN NOESY and aromatic NOESY were re-picked and the new NOESY peak lists and new assignment gave closed structure as CS-Rosetta structure, by CYANA or ASDP. The new peak lists with previous deposited assignment gave relative closed structures, but the hole size is larger than that of 4. AR3436A report updated (continued)

9. Besides some missing weak peaks that is reasonable, some strong peak are also unexpectedly missing in both peak lists HarvestCB peak list vs casdNMR peak list in 15Nnoesy HarvestDB vs. casdNMR HarvestDB vs. casdNMR H18 NH W47 HE1

10. In Sparky, export peak lists in xeasy format by “xe”, one should check “includes unassigned peaks” and “include unintegrated peaks” Lists in HarvestCB casdNMR may exclude unassigned peaks

11. Comparing of the “Hole” Size

12. Deposit vs. casdpeak vs. New Old casd peaks with new assignment, the hole became slightly smaller

13. Deposit vs. oldpeakoldshift vs. New Old “complete” July2009 peaks with old assignment, the hole became smaller

14. Deposit vs. 723peaknewshift vs. New Old “complete” July 23th 2009 peaks with new assignment, the hole became smaller

15. Deposit vs. 723peakoldshift vs. New Old “complete” July 23th2009 peaks with new assignment, the hole became almost as small as new peak new assignment (magenta)