Development of NMR based in vitro and in vivo methods to study nucleotide sugar interconversions

Abstract

Polysaccharides are the dominant components of a plant cell wall. Chemically the plant cell wall consists of cellulose microfibrils embedded in a matrix of hemicellulose and pectin. The matrix polysaccharides are made in the Golgi from nucleotide sugars and are then exported to the cell wall. The goal of this thesis is develop in vitro and in vivo NMR methods to study nucleotide sugar interconversions that lead to the starting products for matrix polysaccharide synthesis and to analyze the data using multivariate statistical methods. The in vitro method included the detection of UDP-Apiose formation from UDP-Glucuronic acid in presence of UDP-Apiose/xylose synthase. The organelle method was developed to study the interconversion of nucleotide sugars within the Golgi apparatus.