Crystal structure of a putative glycosylhydrolase (BT_3782) from Bacteroides thetaiotaomicron VPI-5482 at 1.89 A resolution

Joint Center for Structural Genomics (JCSG)

4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID

Flat mirror (vertical focusing); single crystal Si(111) bent monochromator (horizontal focusing)

20.00% polyethylene glycol 10000, 8.00% ethylene glycol, 0.1M HEPES pH 7.5, 0.005M mannan, 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 RESIDUES 25-387 OF THE TARGET SEQUENCE.

1. 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. 2. ATOM RECORD CONTAINS SUM OF TLS AND RESIDUAL B FACTORS. ANISOU RECORD CONTAINS SUM OF TLS AND RESIDUAL U FACTORS. 3. THE MAD PHASES WERE USED AS RESTRAINTS DURING REFINEMENT. 4. NCS RESTRAINTS WERE IMPOSED BY AUTOBUSTER'S LSSR PROCEDURE (-AUTONCS) 5. EDO (ETHYLENE GLYCOL) AND EPE (HEPES) LIGANDS FROM CRYO- AND CRYS CONDITIONS ARE MODELED INT THE STRUCTURE. 6. THERE IS DISORDERED LOOP IN THE MODELED STRUCTURE 67-70.

Glycosyl hydrolase family 76, PF03663 family, Structural Genomics, Joint Center for Structural Genomics, JCSG, Protein Structure Initiative, PSI-BIOLOGY, HYDROLASE