5LFG

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


X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
IDMethodpHTemperatureDetails
1MICRODIALYSIS8277Crystallization 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).
Crystal Properties
Matthews coefficientSolvent content
3.1661.06

Crystal Data

Unit Cell
Length ( Å )Angle ( ˚ )
a = 86.206α = 90
b = 87.256β = 101.81
c = 109.648γ = 90
Symmetry
Space GroupC 1 2 1

Diffraction

Diffraction Experiment
ID #Crystal IDScattering TypeData Collection TemperatureDetectorDetector TypeDetailsCollection DateMonochromatorProtocol
11x-ray100CCDENRAF-NONIUS2000-07-10MSINGLE WAVELENGTH
Radiation Source
ID #SourceTypeWavelength ListSynchrotron SiteBeamline
1ROTATING ANODERIGAKU MICROMAX-007 HF1.5418

Data Collection

Overall
ID #Resolution (High)Resolution (Low)Percent Possible (Observed)R Merge I (Observed)Net I Over Average Sigma (I)RedundancyNumber Reflections (All)Number Reflections (Observed)Observed Criterion Sigma (F)Observed Criterion Sigma (I)B (Isotropic) From Wilson Plot
11.94893.50.07342.655016
Highest Resolution Shell
ID #Resolution (High)Resolution (Low)Percent Possible (All)Percent Possible (Observed)R Merge I (Observed)Mean I Over Sigma (Observed)RedundancyNumber Unique Reflections (All)
11.9480.40.4

Refinement

Statistics
Diffraction IDStructure Solution MethodCross Validation methodResolution (High)Resolution (Low)Number Reflections (R-Free)Percent Reflections (Observed)R-WorkR-FreeR-Free Selection DetailsMean Isotropic B
X-RAY DIFFRACTIONTHROUGHOUT1.948447993.50.1810.245RANDOM
Temperature Factor Modeling
Anisotropic B[1][1]Anisotropic B[1][2]Anisotropic B[1][3]Anisotropic B[2][2]Anisotropic B[2][3]Anisotropic B[3][3]
Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen AtomsNumber
Protein Atoms4461
Nucleic Acid Atoms
Solvent Atoms213
Heterogen Atoms180

Software

Software
Software NamePurpose
SHELXLrefinement
DENZOdata reduction
SCALEPACKdata scaling
AMoREphasing