Crystal structure of putative glucoamylase (YP_210071.1) from Bacteroides fragilis NCTC 9343 at 2.12 A resolution

Joint Center for Structural Genomics (JCSG)

Flat collimating mirror, toroid focusing mirror

15.8% polyethylene glycol 3350, 0.1M potassium fluoride, 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. THE CLONED CONSTRUCT CONTAINS RESIDUES 23-451 OF THE FULL LENGTH PROTEIN.

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 RESIDUAL B FACTORS ONLY.
4. POTASSIUM, ETHYLENE GLYCOL AND CHLORIDE,
PRESENT IN CRYSTALLIZATION/CRYO CONDITIONS, WERE
MODELED INTO THE STRUCTURE.
5. THE CIS PEPTIDES BETWEEN 222-223 ARE SUPPORTED BY
WELL DEFINED DENSITY.

YP_210071.1, putative glucoamylase, Structural Genomics, Joint Center for Structural Genomics, JCSG, Protein Structure Initiative, PSI-2, Unknown function, HYDROLASE