1E6N

Chitinase B from Serratia marcescens inactive mutant E144Q in complex with N-acetylglucosamine-pentamer

Summary for 1E6N

• Entry DOI: 10.2210/pdb1e6n/pdb
• Descriptor: CHITINASE B, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, SULFATE ION, ... (5 entities in total)
• Functional Keywords: chitin degradation, hydrolase, glycosidase
• Biological source: SERRATIA MARCESCENS
• Total number of polymer chains: 2
• Total formula weight: 114346.86

Authors

Komander, D.,Synstad, B.,Eijsink, V.G.H.,Van Aalten, D.M.F. (deposition date: 2000-08-21, release date: 2001-06-22, Last modification date: 2024-10-16)

Primary citation

Van Aalten, D.M.F.,Komander, D.,Synstad, B.,Gseidnes, S.,Peter, M.G.,Eijsink, V.G.H.
Structural Insights Into the Catalytic Mechanism of a Family 18 Exo-Chitinase
Proc.Natl.Acad.Sci.USA, 98:8979-, 2001

PubMed Abstract: Chitinase B (ChiB) from Serratia marcescens is a family 18 exo-chitinase whose catalytic domain has a TIM-barrel fold with a tunnel-shaped active site. We have solved structures of three ChiB complexes that reveal details of substrate binding, substrate-assisted catalysis, and product displacement. The structure of an inactive ChiB mutant (E144Q) complexed with a pentameric substrate (binding in subsites -2 to +3) shows closure of the "roof" of the active site tunnel. It also shows that the sugar in the -1 position is distorted to a boat conformation, thus providing structural evidence in support of a previously proposed catalytic mechanism. The structures of the active enzyme complexed to allosamidin (an analogue of a proposed reaction intermediate) and of the active enzyme soaked with pentameric substrate show events after cleavage of the glycosidic bond. The latter structure shows reopening of the roof of the active site tunnel and enzyme-assisted product displacement in the +1 and +2 sites, allowing a water molecule to approach the reaction center. Catalysis is accompanied by correlated structural changes in the core of the TIM barrel that involve conserved polar residues whose functions were hitherto unknown. These changes simultaneously contribute to stabilization of the reaction intermediate and alternation of the pKa of the catalytic acid during the catalytic cycle.

PubMed: 11481469
DOI: 10.1073/PNAS.151103798

Experimental method

X-RAY DIFFRACTION (2.25 Å)