3MAT

E.COLI METHIONINE AMINOPEPTIDASE TRANSITION-STATE INHIBITOR COMPLEX

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	298
Detector technology	IMAGE PLATE
Collection date	1998-05-21
Detector	RIGAKU RAXIS IV
Spacegroup name	P 1 21 1
Unit cell lengths	39.297, 65.155, 51.371
Unit cell angles	90.00, 106.12, 90.00

Refinement procedure

Resolution	37.800 - 2.100 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2mat
RMSD bond length	0.013
RMSD bond angle	2.300
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	TNT (5F PRERELEASE)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	37.800	2.180
High resolution limit [Å]	2.100	2.100
Rmerge	0.076 *	0.255 *
Total number of observations	45928 *
Number of reflections	13762
<I/σ(I)>	20.5	4.9
Completeness [%]	94.0	92.9
Redundancy	3.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, sitting drop *	6.1		CRYSTALS OF THE CO(II)-SUBSTITUTED ENZYME WERE GROWN AT ROOM TEMPERATURE BY VAPOR DIFFUSION IN 20-30 UL SITTING DROPS AFTER MIXING THE PROTEIN, 12 MG/ML SOLUTION IN STORAGE BUFFER(25 MM HEPES PH 6.8, 25 MM K2SO4, 100 MM NACL, 1 MM COCL2, 15 MM METHIONINE),CONTAINING 48.8 MM N-OCTANOYL SUCROSE, 1:1 WITH WELL SOLUTIONS (24-26% PEG4000, 0.1M HEPES PH7.0-7.2,FRESH 2 MM COCL2). CRYSTALS WERE OBTAINED OF THE INHIBITOR COMPLEX BY INCUBATING THE PROTEIN AS ABOVE AT ROOM TEMPERATURE FOR 5 MIN WITH A 20-FOLD MOLAR EXCESS OF THE INHIBITOR DISSOLVED IN DMSO. THE FINAL INHIBITOR:ENZYME RATIO WAS 10:1 (1% DMSO) AFTER MIXING THE PREFORMED COMPLEX WITH WELL SOLUTION (0.1M MES PH 6.1,

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir	MES	0.1 (M)
2	1	reservoir		2 (mM)
3	1	reservoir	PEG4000	27-31 (%)
4	1	drop	protein	12 (mg/ml)
5	1	drop	N-octanoyl sucrose	48.8 (mM)