AGAO covalent complex with Benzylhydrazine

Complexes of the Copper-Containing Amine Oxidase from Arthrobacter Globiformis with the Inhibitors Benzylhydrazine and Tranylcypromine.

RESIDUE A382 WAS AN ACTIVE SITE TYROSINE RESIDUE, WHICH WAS AUTOCATALYTICALLY MODIFIED TO BECOME TPQ, AND WAS THEN COVALENTLY LINKED TO THE SUICIDE INHIBITOR BENZYLHYDRAZINE.

ARTHROBACTER GLOBIFORMIS COPPER AMINE OXIDASE, AMINE OXIDASE

AGAO CRYSTALS WERE GROWN IN ABOUT 2 WEEKS BY HANGING-DROP VAPOR DIFFUSION AT 293K. THE RESERVOIR WAS 1.0-1.5M (NH4)2SO4, 2MM CUSO4, 0.15M NA CITRATE, PH 6.0-7.5. THE DROPS WERE 1.5MICROL OF 11.7 MG/ML PROTEIN, 0.05M HEPES, PH 7.0, PLUS 1.5MICROL OF RESERVOIR SOLUTION. A CRYSTAL WAS CRYOPROTECTED BY GRADUAL INCREMENTAL SOAKING IN RESERVOIR SOLUTION MIXED WITH GLYCEROL. THE CONCENTRATION OF GLYCEROL WAS INCREASED FROM 0 TO 30% IN 2-3% INCREMENTS DURING 2 HOURS. FOLLOWING CRYOPROTECTION, THE CRYSTAL WAS SOAKED FOR 15 MINUTES IN CRYOPROTECTANT WITH THE ADDITION OF 2 MILLIMOLAR BENZYLHYDRAZINE DIHYDROCHLORIDE (SIGMA). THE CRYSTAL WAS THEN FROZEN IN THE CRYOSTREAM.

AGAO HOLOENZYME IN A SMALL CELL, AT 2.2 ANGSTROMS

SHEET
THE SHEET STRUCTURE OF THIS MOLECULE IS BIFURCATED. IN
ORDER TO REPRESENT THIS FEATURE IN THE SHEET RECORDS BELOW,
TWO SHEETS ARE DEFINED.

CATALYTIC ACTIVITY: RCH(2)NH(2) + H(2)O + O(2) = RCHO + NH(3) + H(2)O(2).REMARK 500
RESIDES 1-2, MT, ARE THOUGHT TO BE CLEAVED. THE WILD TYPE MATURE PROTEIN IS COMPOSED OF RESIDUES 3-638. RESIDUES 639- 640, SN, IS A PRE-TAG SPACER. RESIDUES 641-648, WSHPQFEK, IS A STREP-TAG II. SEE JUDA ET AL. (2001) PROTEIN EXPRESSION AND PURIFICATION, 22, 455-461

HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 3TY WAS INITIALLY REFINED WITH MINIMAL RESTRAINTS.IT WAS DETERMINED THAT THE 3TY WAS IN THE SUBSTRATE SCHIFF BASE FORM. IN LATE REFINEMENT, THE 3TY WAS RESTRAINED TO THE SUBSTRATE SCHIFF BASE FORM.

AMINE OXIDASE, ARTHROBACTER GLOBIFORMIS, COPPER CONTAINING, METAL-BINDING, OXIDOREDUCTASE, TPQ, QUINONE, INHIBITED, BH, BENZYLHYDRAZINE, 3TY