Crystal structure of a Putative aldose 1-epimerase (b2544) from ESCHERICHIA COLI K12 at 1.59 A resolution

Joint Center for Structural Genomics (JCSG)

Flat collimating mirror, toroid focusing mirror

G(MSE)TIYTLSHGSLKLDVSDQGGVIEGFWRDTTPLLRPGKKSGVATDASCFPLVPFANRVSGNRFVWQGREYQLQPNV
EWDAHYLHGDGWLGEWQCVSHSDDSLCLVYEHRSGVYHYRVSQAFHLTADTLTVTLSVTNQGAETLPFGTGWHPYFPLSP
QTRIQAQASGYWLEREQWLAGEFCEQLPQELDFNQPAPLPRQWVNNGFAGWNGQARIEQPQEGYAII(MSE)ETTPPAPC
YFIFVSDPAFDKGYAFDFFCLEP(MSE)SHAPDDHHRPEGGDLIALAPGESTTSE(MSE)SLRVEWL

GMTIYTLSHGSLKLDVSDQGGVIEGFWRDTTPLLRPGKKSGVATDASCFPLVPFANRVSGNRFVWQGREYQLQPNVEWDA
HYLHGDGWLGEWQCVSHSDDSLCLVYEHRSGVYHYRVSQAFHLTADTLTVTLSVTNQGAETLPFGTGWHPYFPLSPQTRI
QAQASGYWLEREQWLAGEFCEQLPQELDFNQPAPLPRQWVNNGFAGWNGQARIEQPQEGYAIIMETTPPAPCYFIFVSDP
AFDKGYAFDFFCLEPMSHAPDDHHRPEGGDLIALAPGESTTSEMSLRVEWL

DATA WERE SCALED USING XSCALE WITH FRIEDEL PAIRS KEPT AS SEPARATE WHEN COMPUTING R-SYM, COMPLETENESS AND <I/SIGMA(I)>

0.2000M magnesium chloride, 20.0000% polyethylene glycol 8000, 0.1M TRIS pH 8.5, Additive 0.005M D-GLUCOSE, NANODROP', VAPOR DIFFUSION, SITTING DROP, temperature 277K

THIS 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.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 FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 3.ATOM RECORDS CONTAIN SUM OF TLS AND RESIDUAL B FACTORS. ANISOU RECORD CONTAINS SUM OF TLS AND RESIDUAL U FACTORS. 4.SOLVENT MOLECULES WERE EXCLUDED FROM AUTOMATIC TLS ASSIGNMENT. 5.PEG-200 AND PEG-8000 FRAGMENTS (PEG) AND MAGNESIUM (MG) FROM THE CRYSTALLIZATION AND CRYOPROTECTANT SOLUTIONS HAVE BEEN MODELED.

CRYSTAL PACKING ANALYSIS AND ANALYTICAL SIZE-EXCLUSION CHROMATOGRAPHY SUGGEST THE ASSIGNMENT OF A MONOMER AS THE SIGNIFICANT OLIGOMERIZATION STATE IN SOLUTION.

Structural Genomics, Joint Center for Structural Genomics, JCSG, Protein Structure Initiative, PSI-2, ISOMERASE

MTIYTLSHGSLKLDVSDQGGVIEGFWRDTTPLLRPGKKSGVATDASCFPLVPFANRVSGNRFVWQGREYQLQPNVEWDAH
YLHGDGWLGEWQCVSHSDDSLCLVYEHRSGVYHYRVSQAFHLTADTLTVTLSVTNQGAETLPFGTGWHPYFPLSPQTRIQ
AQASGYWLEREQWLAGEFCEQLPQELDFNQPAPLPRQWVNNGFAGWNGQARIEQPQEGYAIIMETTPPAPCYFIFVSDPA
FDKGYAFDFFCLEPMSHAPDDHHRPEGGDLIALAPGESTTSEMSLRVEWL