4S0Q

The X-ray structure of the adduct formed in the reaction between bovine pancreatic ribonuclease and carboplatin

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	100.521, 32.499, 72.711
Unit cell angles	90.00, 89.59, 90.00

Refinement procedure

Resolution	72.710 - 2.090
R-factor	0.20801
Rwork	0.205
R-free	0.26175
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	pdb code 1JVT chain A without water molecules and ligands
RMSD bond length	0.016
RMSD bond angle	1.762
Refinement software	REFMAC (5.8.0049)

Data quality characteristics

	Overall
Low resolution limit [Å]	72.710
High resolution limit [Å]	2.090
Rmerge	0.124
Number of reflections	14095
Completeness [%]	99.7
Redundancy	5.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5	298	Crystals of the RNase A-carboplatin adduct have been obtained by soaking experiments where pre-grown monoclinic protein crystals were incubated with an excess of the platinum drug (at a protein to platinum ratio 1:10). Crystals of RNase A were grown by hanging-drop vapor mixing 1 L of RNase A at 20 mg mL-1 with equal volumes of reservoir solution containing 20% PEG4000 and 20 mM sodium citrate buffer pH 5.0 at 298 K. Two weeks after their appearance, crystals were soaked for four days in a solution of Carboplatin dissolved in 10 L of reservoir. At the final, Pt drug:protein concentrations were in 10:1 ratio., VAPOR DIFFUSION, HANGING DROP