[sharp-discuss] H32 SHARP-ARP/wARP-Refmac problem?

Ole Kristensen OleK@sc.dfh.dk
Tue, 10 Dec 2002 11:22:04 +0100


In a current case, I got so far as to run the arp/warp step from autosharp
(SHARP 2.0.0, Sushi 3.0.14). Despite good phases I got stuck. Here is the
log output:
MTZ file	:
/home/sharp/users/olek/None.sharp/logfiles/tig118.5/eden_flat_34.5pc.mtz

$warpbin	:	/a/xtal/linux/arp_warp_6.0/bin/Linux	
Cell	:	71.6440 71.6440 375.4090 90.0000 90.0000 120.0000	
Spacegroup	:	H32 (155)	
Wilson B-Factor	:	38.419	
Protein size	:	2952	
Sequence file	:	<wARP_34.5pc/seq.pir>	
Resolution	:	30.000 2.30	
Asymmetric unit	:	0.00000 1.00000 0.00000 1.00000 0.00000 1.00000	
2. Generating "warp.par" for ARP/wARP version 6.0 
(details) <wARP_34.5pc/warp.par> (directory index) <wARP_34.5pc> 
3. Running "arpwarp.sh"
1. reading warp.par (for ARP/wARP version 6.0)
2. Building free atoms model
2.1 calculating initial map
2.2 expanding map to ARP limits
2.3 building initial model
Note: Had to go as low as 0.80 sigma, to complete atoms search.
 
Note: Remove atoms if below 1.0 sigmas in 2mFoDFc map.
      Add atoms in density above 3.2 sigmas in mFoDFc map.
      A maximum of 100 ARP refinement cycles will be run in total.
      Rebuilding of the model will happen every 10 cycles.
3. running warpNtrace loop
 
Note: using fixed weight in REFMAC of 0.5 - check geometry afterwards!
 
 
Note: automatically adjusting phase blurring in REFMAC
 
  16247 reflections (   99.99 % complete ) and 0 restraint used, for
refining 3310 atoms.
  Observations/parameters ratio is 1.23 (~1.23 with restraints).

  Starting model : R = 0.341 (Rfree = 0.331)

  Cycle   1 : After refmac :   R = 0.314 (Rfree = 0.320) 
              Found 21 (21 requested) and removed 15 (16 requested) atoms.
  Cycle   2 : After refmac :   R = 0.297 (Rfree = 0.321) 
              Found 21 (21 requested) and removed 2 (16 requested) atoms.
  Cycle   3 : After refmac :   R = 0.286 (Rfree = 0.324) 
              Found 21 (21 requested) and removed 5 (18 requested) atoms.
  Cycle   4 : After refmac :   R = 0.278 (Rfree = 0.322) 
              Found 22 (22 requested) and removed 9 (21 requested) atoms.
  Cycle   5 : After refmac :   R = 0.271 (Rfree = 0.321) 
              Found 22 (22 requested) and removed 10 (22 requested) atoms.
  Cycle   6 : After refmac :   R = 0.265 (Rfree = 0.317) 
              Found 22 (22 requested) and removed 13 (22 requested) atoms.
  Cycle   7 : After refmac :   R = 0.260 (Rfree = 0.315) 
              Found 22 (22 requested) and removed 20 (22 requested) atoms.
  Cycle   8 : After refmac :   R = 0.255 (Rfree = 0.312) 
              Found 22 (22 requested) and removed 12 (22 requested) atoms.
  Cycle   9 : After refmac :   R = 0.253 (Rfree = 0.313) 
              Found 22 (22 requested) and removed 21 (22 requested) atoms.
  Cycle  10 : After refmac :   R = 0.249 (Rfree = 0.309) 
              Found 22 (22 requested) and removed 7 (22 requested) atoms.

  Building Cycle 1
   Round 1 : 291 peptides in 22 chains.
   Round 2 : 312 peptides in 14 chains.
     Chains 14, Residues 298 [out of 369], Connectivity Index 0.91

  Cycle  11 : After refmac :   R = 0.339 (Rfree = 0.384) 
              Found 16 (16 requested) and removed 8 (8 requested) atoms.
  Cycle  12 : After refmac :   R = 0.317 (Rfree = 0.375) 
              Found 16 (16 requested) and removed 8 (8 requested) atoms.
  Cycle  13 : After refmac :   R = 0.303 (Rfree = 0.366) 
              Found 16 (16 requested) and removed 8 (8 requested) atoms.
  Cycle  14 : After refmac :   R = 0.293 (Rfree = 0.359) 
              Found 16 (16 requested) and removed 8 (8 requested) atoms.
  Cycle  15 : After refmac :   R = 0.283 (Rfree = 0.348) 
              Found 16 (16 requested) and removed 8 (8 requested) atoms.
  Cycle  16 : After refmac :   R = 0.277 (Rfree = 0.345) 
              Found 16 (16 requested) and removed 8 (8 requested) atoms.
  Cycle  17 : After refmac :   R = 0.270 (Rfree = 0.323) 
              Found 17 (17 requested) and removed 8 (8 requested) atoms.
  Cycle  18 : After refmac :   R = 0.263 (Rfree = 0.321) 
              Found 17 (17 requested) and removed 8 (8 requested) atoms.
  Cycle  19 : After refmac :   R = 0.258 (Rfree = 0.317) 
              Found 17 (17 requested) and removed 8 (8 requested) atoms.
  Cycle  20 : After refmac :   R = 0.251 (Rfree = 0.316) 
              Found 17 (17 requested) and removed 8 (8 requested) atoms.

  Building Cycle 2
   Round 1 : 285 peptides in 14 chains.
   Round 2 : 307 peptides in 13 chains.
     Chains 10, Residues 285 [out of 369], Connectivity Index 0.92
 
Note: had to fix PDB file "./files/reshuffle_cycle20.pdb" for RES. numbering
- please check programs!
 
ERROR : program terminated in REFMAC5 (see refmac_cycle21.log
<wARP_34.5pc/refmac_cycle21.log>)

The ./files/reshuffle_cycle20.pdb looks okay. Refmac seems to be the
problem. I tried
both the refmac5 version from ccp4-4.2.2 and garib's latest version. Here,
is a bit
of my latest attempt:
###############################################################
 ###############################################################
 ###############################################################
 ### CCP4 4.2: Refmac_5.2.00      version 5.2.00    : 31/10/01##
 ###############################################################
 User:   Run date:  9/12/02  Run time:23:43:19
......
Data line--- MAKE HYDR N
 Data line--- MAKE CHECK 0
 Data line--- LABI FP=FPsha SIGFP=SIGFPsha FREE=FreeR_flag HLA=HLAdm
HLB=HLBdm HLC=HLCdm HLD=HLDdm
 Data line--- LABO FC=FC PHIC=PHIC      FWT=2FOFCWT      PHWT=PH2FOFCWT
DELFWT=FOFCWT PHDELWT=PHFOFCWT
 Data line--- REFI RESOLUTION 30.000 2.30 TYPE RESTR
 Data line--- REFI RESI MLKF PHASed SCBLurred 0.34
 Data line--- REFI BREF ISOTROPIC METH CGMAT
 Data line--- WEIG MATRIX 0.5
 Data line--- DAMP 1.0 1.0
 Data line--- SCAL TYPE  BULK LSSC ANIS  FIXBulk
 Data line--- SCAL MLSC  FIXBulk
 Comment line--- ! maybe only at later stages?
 Data line--- SOLVENT YES
 Data line--- NCYC 1
 Data line--- MONI MANY
 Data line--- BLIMI 2 80
 Data line--- BINS 20
 Data line--- VDWR SIGMA DUMMY 50.0 INCREMENT DUMMY -1.0
 Data line--- VDWR 0.0
 Data line--- END
.....
.....
     CGMAT cycle number =      1
Too many atoms in one brick.
Maximum allowed in one brick =         60
Check symmetry, cell dimensions
===> Error: CREATE_NET failed
 Refmac_5.2.00:  CREATE_NET failed
 Times: User:       5.0s System:    0.7s Elapsed:    0:13

Any ideas about how to get around this? I was wondering if it is related to
the asymmetric unit being defined as:
Asymmetric unit	:	0.00000 1.00000 0.00000 1.00000 0.00000 1.00000	

Clemens has previously pointed out that one could possibly change the
settings in
$BDG_home/database/lists/symm.dat. Will it work to change the field 10 for
spacegroup
H32 to something smaller than the 0 1 0 1 0 1 limits distributed?
When I simply take the starting file eden_flat_34.5pc.mtz and run arpwarp
just from the
ccp4i with default settings a smaller asu seems to be used and thing run
without error.


Best regards,
Ole Kristensen
Denmark