1AP2

SINGLE CHAIN FV OF C219

Experimental procedure

Source type	ROTATING ANODE
Source details	RIGAKU
Temperature [K]	298
Detector technology	AREA DETECTOR
Collection date	1997-05
Detector	XUONG-HAMLIN MULTIWIRE
Spacegroup name	P 21 21 21
Unit cell lengths	59.040, 64.350, 154.110
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	6.000 - 2.360
R-factor	0.206 *
Rwork	0.204
R-free	0.28500 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1igm
RMSD bond length	0.006
RMSD bond angle	1.110 *
Data reduction software	SDMS
Data scaling software	SDMS
Phasing software	X-PLOR (3.851)
Refinement software	X-PLOR (3.851)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.550
High resolution limit [Å]	2.360	2.360
Rmerge	0.098 *	0.277 *
Total number of observations	81247 *
Number of reflections	22616	3671 *
<I/σ(I)>	9.7	2.8
Completeness [%]	91.6	76.1
Redundancy	3.6	2.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.5		THE PROTEIN WAS CRYSTALLIZED IN 21% PEG 6000, 100 MM SODIUM CITRATE PH 4.5, IN HANGING DROPS CONTAINING SUBTILISIN CARLSBERG IN A 1:100 MOLAR RATIO., vapor diffusion - hanging drop THE DEPOSITORS CRYSTALLIZED THE SINGLE CHAIN FV IN THE PRESENCE OF SUBTILISIN, AS DESCRIBED IN THE PRIMARY REFERENCE. THE SEQUENCE OF THE SINGLE CHAIN IS DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWAS TRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEP (LIGHT CHAINS A AND C) GGGGSGGGGSGKSGGGG (LINKER) EVQLQQSGAELVRPGASVKLSCTASGFNIKDDFMHWVKQRPEQGLEWIGRIDPANDNT KYAPKFQDKATIIADTSSNTAYLQLSSLTSEDTAVYYCARREVYSYYSPLDVWGAGTT VTVPSG (HEAVY CHAINS B AND D) SEQKLISEEDLNHHHHH (C-MYC TAG + 5XHIS TAG) THE DEPOSITORS HAVE NOT DETERMINED WHERE SUBTILISIN CLEAVES EXACTLY, BUT THE MOBILITY ON SDS GELS SUGGESTS THAT THE MOST OF THE LINKER AND MOST OF BOTH TAGS ARE REMOVED.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	7 (mg/ml)
2	1	drop	Tris-HCl	10 (mM)
3	1	reservoir	sodium citrate	100 (mM)
4	1	reservoir	PEG6000	21 (%(w/v))
5	1	reservoir	subtilisin