Sde 2656
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Data Integration & Reduction
- Use CCP4i & Mosflm
MR (Phaser)
- get MR models (PDB homologs) - 1TZ6_A, 2V78_A, 1TYY_A, 2DCN_A, 3IQ0_A, 1V19_A (Top 6 PDBs) by Psi-blast
- superimpose them into 1TZ6_A (make multiple models & compare those structures) by UCSF chimera
Phenix.automr
- 1st trial: use multiple models
- it didn't give any good solution
- 2nd trial: try each model looking for the best solution
- 1V19A gave best solution
- resolution 4-20 was enough to find the solution
- judgement: LLG>100, Z-score>7
- our solution was almost at edge
Solution annotation (history): RFZ=5.2 TFZ=4.6 PAK=0 LLG=35 RFZ=5.6 TFZ=7.7 PAK=1 LLG=92 LLG=92 Log likelihood gain for solution MR: 92.0433
- Thankabouts
- how about using HM (homology model)?
- superimpose HM into best solution
- use C-alpha only model
- try Autobuild with these models
Phenix.autobuild
- Going on to AutoBuild with this solution (it took about 24 hrs)
phenix.autobuild \ data=AutoBuild_run_1_/exptl_fobs_phases_freeR_flags.mtz \ model=AutoBuild_run_1_/cycle_best_9.pdb \ rebuild_in_place=True \ seq_file=sde2656.seq SOLUTION CYCLE R RFREE BUILT PLACED 1 2 0.37 0.44 575 328 2 3 0.32 0.40 503 235 3 4 0.28 0.35 515 359 4 5 0.27 0.34 529 388 5 6 0.26 0.33 545 400 6 7 0.25 0.31 551 407 7 8 0.25 0.31 555 411 8 9 0.24 0.30 552 446
- The model was partial (~50 residues in C-term missing)