Comparative subcellular immunolocation of polypeptides associated with xylan and callose synthases in French bean (Phaseolus vulgaris) during secondary wall formation.
The Golgi apparatus of plant cells is thought to be the main site of synthesis of cell wall matrix polysaccharides and the terminal glycosylation of glycoproteins. Much of this evidence still depends on earlier biochemical studies employing subcellular fractionation. However acquiring pure Golgi membranes is still difficult and the question of spatial organisation of glycosyl transferases can be addressed by immunolocation of the enzymes. An antibody to a xylan synthase-associated polypeptide from French bean, the enzyme which synthesises the core polysaccharide for secondary wall xylan, has been raised and shown to inhibit its activity. Xylan is deposited in secondary thickenings and the xylan synthase was only detected in appreciable amounts in developing xylem cells. The location within the Golgi stack was observed throughout the dictyosomes. Some enzyme subunits were also detected in post-Golgi vesicles. A second antibody to a non-catalytic M(r) 65000 subunit of beta 1,3- glucan (callose) synthase was used for a comparative study. Although the bulk of this enzyme has been detected in previous studies at plasmamembrane-wall interfaces in sieve plates and stressed tissue, a Golgi-location can be observed in root tip meristematic cells during cell plate formation. The enzyme was present throughout the stacks. Callose was also immunolocated in a similar manner to xylan in secondary walls and thickenings and in pits in developing xylem. In these cells, the callose synthase was detected at the surface of the growing thickenings and the plasmamembrane within the pits.
Gregory
Abigail C E
AC
School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, UK.
Smith
Colin
C
Kerry
Maria E
ME
Wheatley
Edward R
ER
Bolwell
G Paul
GP
D015153:Blotting, Western; D002473:Cell Wall; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005964:Glucosyltransferases / Q000378:metabolism*; D008565:Membrane Proteins; D010430:Pentosyltransferases / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D027805:Phaseolus / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins; D013347:Subcellular Fractions / Q000378:metabolism*
D008565:Membrane Proteins; D010455:Peptides; D029702:Schizosaccharomyces pombe Proteins; D005964:Glucosyltransferases; C020570:1,3-beta-glucan synthase; D010430:Pentosyltransferases; C038331:1,4-beta-D-xylan synthase