TY - JOUR
T1 - Dynamic elements govern the catalytic activity of CapE, a capsular polysaccharide-synthesizing enzyme from Staphylococcus aureus
AU - Miyafusa, Takamitsu
AU - Caaveiro, Jose M.M.
AU - Tanaka, Yoshikazu
AU - Tsumoto, Kouhei
N1 - Funding Information:
This work was supported in part by a Grant-in-Aid for General Research from the Japan Society for the Promotion of Science (to K.T.). The funding agency had no role in the design of the study, the collection, analysis and interpretation of the data, the writing of the report, and in the decision to submit the manuscript for publication.
PY - 2013/11/29
Y1 - 2013/11/29
N2 - CapE is an essential enzyme for the synthesis of capsular polysaccharide (CP) of pathogenic strains of Staphylococcus aureus. Herein we demonstrate that CapE is a 5-inverting 4,6-dehydratase enzyme. However, in the absence of downstream enzymes, CapE catalyzes an additional reaction (5-back-epimerization) affording a by-product under thermodynamic control. Single-crystal X-ray crystallography was employed to identify the structure of the by-product. The structural analysis reveals a network of coordinated motions away from the active site governing the enzymatic activity of CapE. A second dynamic element (the latch) regulates the enzymatic chemoselectivity. The validity of these mechanisms was evaluated by site-directed mutagenesis.
AB - CapE is an essential enzyme for the synthesis of capsular polysaccharide (CP) of pathogenic strains of Staphylococcus aureus. Herein we demonstrate that CapE is a 5-inverting 4,6-dehydratase enzyme. However, in the absence of downstream enzymes, CapE catalyzes an additional reaction (5-back-epimerization) affording a by-product under thermodynamic control. Single-crystal X-ray crystallography was employed to identify the structure of the by-product. The structural analysis reveals a network of coordinated motions away from the active site governing the enzymatic activity of CapE. A second dynamic element (the latch) regulates the enzymatic chemoselectivity. The validity of these mechanisms was evaluated by site-directed mutagenesis.
KW - Capsular polysaccharide
KW - Conformational change
KW - Pathogenic bacterium
KW - SDR enzyme
KW - Staphylococcus aureus
KW - UDP-sugar
KW - X-ray crystallography
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U2 - 10.1016/j.febslet.2013.10.009
DO - 10.1016/j.febslet.2013.10.009
M3 - Article
C2 - 24157361
AN - SCOPUS:84888001994
VL - 587
SP - 3824
EP - 3830
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 23
ER -