Crystal structure of Zn-containing NagZ H174A mutant from Pseudomonas aeruginosa

Catalytic Cycle of the N-Acetylglucosaminidase NagZ from Pseudomonas aeruginosa.

BOTH PROTEIN CHAINS CONTAIN A HIS RESIDUE AT POSITION -1 FROM THE FUSION TAG USED FOR PURIFICATION. THE SEQUENCE INCLUDES THE SINGLE-POINT MUTATION H174A THAT MAKES THIS PROTEIN INACTIVE.

BETA-N-ACETYLHEXOSAMINIDASE, N-ACETYL-BETA-GLUCOSAMINIDASE, NAGZ

MGSSHHHHHHSSGLVPRGSHMQGSLMLDIGGTWLTAEDRQILRHPEVGGLIIFARNIEHPAQVRELCAAIRAIRPDLLLA
VDQEGGRVQRLRQGFVRLPAMRAIADNPNAEELAEHCGWLMATEVQAVGLDLSFAPVLDLDHQRSAVVGSRAFEGDPERA
ALLAGAFIRGMHAAGMAATGKHFPGHGWAEADSAVAIPEDARSLEEIRRSDLVPFARLAGQLDALMPAHVIYPQVDPQPA
GFSRRWLQEILRGELKFDGVIFSDDLSMAGAHVVGDAASRIEAALAAGCDMGLVCNDRASAELALAALQRLKVTPPSRLQ
RMRGKGYANTDYRQQPRWLEALSALRAAQLID

30% PEG 8000 100 MM SODIUM CACODYLATE PH 6.0 200 MM SODIUM ACETATE PH 6.0

ZINC ION (ZN): THE SOURCE OF ZN IS UNKNOWN. IT IS LIKELY
THAT ZN COMES EITHER THE EXPRESSION HOST OR THE
PURIFICATION BUFFERS
DI(HYDROXYETHYL)ETHER (PEG): PEG IS PRESENT IN THE
CRYSTALLIZATION CONDITION

THE EXPERIMENTAL SEQUENCE CONTAINS A HIS-TAG AT THE N- TERMINUS. THERE IS ALSO A SINGLE-POINT MUTATION H174A

APO STRUCTURE OF NAGZ FROM PSEUDOMONAS AERUGINOSA

RESIDUES 124-135 AND 171-174 IN CHAIN A, AND REIDUES 124-134, 171-175 AND 249-251 IN CHAIN B WERE NOT MODELED DUE TO POOR ELECTRON DENSITY.

HYDROLASE, CELL-WALL RECYCLING, ANTIBIOTIC RESISTANCE, GLYCOSIDE HYDROLASE, N-ACETYLGLUCOSAMINIDASE, BETA-HEXOSAMINIDASE, PEPTIDOGLYCAN