TY  - JOUR
A1  - Eijsink, Vincent G. H.
A1  - Synstad, Bjoenar
A1  - Gaseidnes, Sigrid
A1  - Komander, David
A1  - Houston, Douglas R.
A1  - Peter, Martin G.
A1  - van Aalten, Daan M. F.
T1  - Structure and function of chitinolytic enzymes
N2  - The recent work on a variety of family 18 chitonolytic enzymes has yielded important data concerning the structure, substrate-binding, catalysis, inhibitor-binding and even dynamics. These data have been useful in helping to better understand the roles of various types of chitinases in chitin hydrolysis, to rationally engineer the properties of these enzymes, thus making them more suitable as biocatalysts, and to study and understand the effectiveness of natural and designed chitinase inhibitors, which may be of medical interest. On the other hand, the recent work on ChiB shows that catalysis in family 18 chitinases is a highly complicated process, involving larger parts of the enzyme and dynamics. Thus, despite recent discoveries, there is still a lot more to discover about how these enzyme work.
Y1  - 2003
SN  - 82-471-5901-5
ER  - 
TY  - JOUR
A1  - van Aalten, Daan M. F.
A1  - Komander, David
A1  - Synstad, Bjoenar
A1  - Gaseidnes, Sigrid
A1  - Peter, Martin G.
A1  - Eijsink, Vincent G. H.
T1  - Structural Insights into the catalytic mechanism of a family 18 exochitinase
N2  - Chitinase B (ChiB) from Serratia marcescens is a family 18 exochitinase 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, substrateassisted catalysis, and product displacement. The structure of an inactive ChiB mutant (E144Q) complexed with a pentameric substrate (binding in subsites 22 to 13) shows closure of the ''roof'' of the active site tunnel. It also shows that the sugar in the 21 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 11 and 12 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.
Y1  - 2002
ER  -