High-Voltage Paper Electrophoresis (HVPE) of Cell-Wall Building Blocks and Their Metabolic Precursors
HVPE is an excellent and often overlooked method for obtaining objective and meaningful information about cell-wall “building blocks” and their metabolic precursors. It provides not only a means of analysis of known compounds but also an insight into the charge and/or mass of any unfamiliar compounds that may be encountered. It can be used preparatively or analytically. It can achieve either “class separations” (e.g. delivering all hexose monophosphates into a single pool) or the resolution of different compounds within a given class (e.g. ADP-Glc from UDP-Glc; or GlcA from GalA). All information from HVPE about charge and mass can be obtained on minute traces of analytes, especially those that have been radiolabelled, e.g. by in-vivo feeding of a 3H- or 14C-labelled precursor. HVPE does not usually damage the substance under investigation (unless staining is used), so samples of interest can be eluted intact from the paper ready for further analysis. Although HVPE is a technique that has been available for several decades, recently it has tended to be sidelined, possibly because the apparatus is not widely available. Interested scientists are invited to contact the author about the possibility of accessing the Edinburgh apparatus.
Key wordsCharge:mass ratio Electrophoresis Hydroxyproline oligoarabinosides Ionisation Monosaccharides Nucleotide-sugars Oligosaccharides Radiolabelling Sugar-phosphates Uronic acids
I thank numerous past and present colleagues and students for electrophoretic data and experience presented in this chapter, and the UK BBSRC for financial support.
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