Crystal structure of a formyltetrahydrofolate deformylase (purU, PP_1943) from PSEUDOMONAS PUTIDA KT2440 at 2.05 A resolution

Joint Center for Structural Genomics (JCSG)

Flat collimating mirror, toroid focusing mirror

G(MSE)NKNNQYVLSLACQDAPGIVSEVSTFLFNNGANIVEAEQFNDEDSSKFF(MSE)RVSVEIPVAGVNDFNSAFGKV
VEKYNAEWWFRPRTDRKKVVI(MSE)VSKFDHCLGDLLYRHRLGELD(MSE)EVVGIISNHPREALSVSLVGDIPFHYLP
VTPATKAAQESQIKNIVTQSQADLIVLARY(MSE)QILSDDLSAFLSGRCINIHHSFLPGFKGAKPYHQAHTRGVKLIGA
TAHFVTADLDEGPIIAQDVEHVSHRDSAEDLVRKGRDIERRVLSRAVLLFLEDRLIVNGERTVVFAD

GMNKNNQYVLSLACQDAPGIVSEVSTFLFNNGANIVEAEQFNDEDSSKFFMRVSVEIPVAGVNDFNSAFGKVVEKYNAEW
WFRPRTDRKKVVIMVSKFDHCLGDLLYRHRLGELDMEVVGIISNHPREALSVSLVGDIPFHYLPVTPATKAAQESQIKNI
VTQSQADLIVLARYMQILSDDLSAFLSGRCINIHHSFLPGFKGAKPYHQAHTRGVKLIGATAHFVTADLDEGPIIAQDVE
HVSHRDSAEDLVRKGRDIERRVLSRAVLLFLEDRLIVNGERTVVFAD

1.0000M NaCitrate, 0.1M CHES pH 9.5, NANODROP, VAPOR DIFFUSION, SITTING DROP, temperature 277K

THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS REMOVED WITH TEV PROTEASE LEAVING ONLY A GLYCINE (0) FOLLOWED BY THE TARGET SEQUENCE.

1.HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2.ATOM RECORD CONTAINS SUM OF TLS AND RESIDUAL B FACTORS. ANISOU RECORD CONTAINS SUM OF TLS AND RESIDUAL U FACTORS. WATERS WERE EXCLUDED FROM AUTOMATIC TLS ASSIGNMENT. 3.A MET-INHIBITION PROTOCOL WAS USED FOR SELENOMETHIONINE INCORPORATION DURING PROTEIN EXPRESSION. THE OCCUPANCY OF THE SE ATOMS IN THE MSE RESIDUES WAS REDUCED TO 0.75 TO ACCOUNT FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 4.AN UNKNOWN LIGAND HAS BEEN MODELED IN THE STRUCTURE. BASED ON DENSITY AND HYDROGEN BONDING DONOR/ACCEPTOR LOCATIONS, THIS MAY BE NITROBENZENE, BENZOATE, OR NICOTINIC ACID. 5. SODIUM (NA) IONS AND CITRATE (FLC) ANIONS FROM THE CRYSTALLIZATION REAGENT, IMIDAZOLE (IMD) FROM THE PROTEIN BUFFER AND ETHYLENE GLYCOL (EDO) FROM THE CRYOPROTECTANT SOLUTION ARE MODELED IN THE STRUCTURE.

ANALYTICAL SIZE EXCLUSION CHROMATOGRAPHY INDICATES A TETRAMER IS A SIGNIFICANT OLIGOMERIZATION STATE.

N-terminal ACT domain, Structural Genomics, Joint Center for Structural Genomics, JCSG, Protein Structure Initiative, PSI-2, HYDROLASE

MNKNNQYVLSLACQDAPGIVSEVSTFLFNNGANIVEAEQFNDEDSSKFFMRVSVEIPVAGVNDFNSAFGKVVEKYNAEWW
FRPRTDRKKVVIMVSKFDHCLGDLLYRHRLGELDMEVVGIISNHPREALSVSLVGDIPFHYLPVTPATKAAQESQIKNIV
TQSQADLIVLARYMQILSDDLSAFLSGRCINIHHSFLPGFKGAKPYHQAHTRGVKLIGATAHFVTADLDEGPIIAQDVEH
VSHRDSAEDLVRKGRDIERRVLSRAVLLFLEDRLIVNGERTVVFAD