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Published on: May 25, 2018
Structure/function analysis of Pasteurella multocida heparosan synthases: toward defining enzyme specificity and
Nigel J Otto1, Dixy E Green, Sayaka Masuko
1Department of Biochemistry and Molecular Biology, Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73126, USA.
Pasteurella multocida heparosan synthases (PmHS1 and PmHS2) create the glycosaminoglycan heparosan. Chimeric enzymes revealed structure-function relationships, identifying regions for donor binding and acceptor usage.
Area of Science:
- Biochemistry
- Glycobiology
- Enzymology
Background:
- Pasteurella multocida Type D harbors homologous bifunctional glycosyltransferases, PmHS1 and PmHS2.
- These enzymes synthesize the glycosaminoglycan heparosan from UDP-glucuronic acid and UDP-N-acetylglucosamine.
Purpose of the Study:
- To investigate the structure-function relationships of Pasteurella multocida heparosan synthases.
- To identify enzyme regions responsible for substrate specificity and catalytic activity.
Main Methods:
- Generation and purification of chimeric heparosan synthase enzymes.
- In vitro radiochemical sugar incorporation assays.
- Comparison of kinetic properties and amino acid sequences.
Main Results:
- Chimeric enzymes demonstrated enzymatic activity, with some producing monodisperse polysaccharides.
- Novel characteristics, including expanded sugar analog usage, were observed.
- Regions influencing donor binding specificity and acceptor usage were identified.
Conclusions:
- Chimeric enzymes provide insights into the structure-function relationships of heparosan synthases.
- Understanding these enzymes aids in comprehending glycosaminoglycan biosynthesis.
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