5LFG

X-ray structure of a new fully ligated carbomonoxy form of Trematomus newnesi hemoglobin (Hb1TnCO).

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	CCD
Collection date	2000-07-10
Detector	ENRAF-NONIUS
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	86.206, 87.256, 109.648
Unit cell angles	90.00, 101.81, 90.00

Refinement procedure

Resolution	8.000 - 1.940
Rwork	0.181
R-free	0.24500
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	SHELXL

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	8.000
High resolution limit [Å]	1.940	1.940
Rmerge	0.073	0.400
Number of reflections	55016
<I/σ(I)>	4
Completeness [%]	93.5	80.4
Redundancy	2.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICRODIALYSIS	8	277	Crystallization trials were performed under CO atmosphere. The dialysis technique was used to obtain protein crystals: the protein, in a 50 mM Tris pH 8.0 buffer with 2mM dithionite, with a concentration of 5 mg x ml-1, was separated by the precipitant reservoir (2.0 M ammonium sulphate, 2 mM dithionite) via a dialysis membrane with a 8000 Da cutoff. Single crystals of the carbomonoxylated Hb1Tn (Hb1TnCO), suitable for X-ray diffraction, were grown in a week (size 0,2 x 0,2 x 0,1 mm3).