2HIS

CELLULOMONAS FIMI XYLANASE/CELLULASE DOUBLE MUTANT E127A/H205N WITH COVALENT CELLOBIOSE

Summary for 2HIS

• Entry DOI: 10.2210/pdb2his/pdb
• Related PRD ID: PRD_900023
• Descriptor: CELLULOMONAS FIMI FAMILY 10 BETA-1,4-GLYCANASE, beta-D-glucopyranose-(1-4)-alpha-D-glucopyranose (3 entities in total)
• Functional Keywords: hydrolase, o-glycosyl, xylanase/cellulase, a/b barrel
• Biological source: Cellulomonas fimi
• Total number of polymer chains: 1
• Total formula weight: 34312.16

Authors

Notenboom, V.,Birsan, C.,Nitz, M.,Rose, D.R.,Warren, R.A.J.,Wither, S.G. (deposition date: 1998-02-23, release date: 1998-10-14, Last modification date: 2024-10-16)

Primary citation

Notenboom, V.,Birsan, C.,Nitz, M.,Rose, D.R.,Warren, R.A.,Withers, S.G.
Insights into transition state stabilization of the beta-1,4-glycosidase Cex by covalent intermediate accumulation in active site mutants.
Nat.Struct.Biol., 5:812-818, 1998

PubMed Abstract: The catalytic mechanism of 'retaining' beta-glycosidases has been the subject of considerable interest and debate for many years. The visualization of a covalent glycosyl enzyme intermediate by X-ray crystallography was first accomplished with a saccharide substrate substituted with fluorine at its 2-position. The structure implicated major roles for residue His 205 and for the 2-hydroxyl position of the proximal saccharide in binding and catalysis. Here we have studied the kinetic behavior of various His 205 mutants. One of these mutants, a double mutant H205N/E127A, has been used to stabilize a covalent glycosyl-enzyme intermediate involving an unsubstituted sugar, permitting crystallographic analysis of the interactions between its 2-hydroxyl group and the enzyme.

PubMed: 9731776
DOI: 10.1038/1852

Experimental method

X-RAY DIFFRACTION (1.84 Å)