title: Purification of several pectin methyltransferases from cell suspension cultures of flax (Linum usitatissimum L.).
abstract: Three pectin methyltransferases (PMT5, PMT7, PMT18; EC 2.1.1.6.x) were solubilized from the endo-membrane complex of flax cells, with 0.05% Triton X-100. After a 3 step-chromatography procedure, PMT7 and PMT5 were purified to apparent homogeneity. PMT5 and PMT7 differed regarding their optimum pH (5 or 7), the methyl acceptor (low or highly methylesterified pectin), their focusing pH range (6-7 or 8-9) and relative molecular mass (40 +/- 5 or 110 +/- 10 kDa). SDS-PAGE of PMT5 and PMT7 did not reveal bands at 40 or 110 kDa but only a silver stained band of about 18 kDa. Two independent methods (photo labelling and enzymatic activity) showed that this silverstained band corresponded to a methyltransferase with affinity for pectins. This polypeptide was of the same size as the enzyme designed PMT18 (18 +/- 3 kDa; pl 4-4.5) recovered during size exclusion chromatography of either PMT7 or PMT5, suggesting that PMT18 bears the catalytic site of PMT5 and PMT7.
authors.affiliation: Scueor Esa 6037 CNRS, université de Rouen, 76821 Mont-Saint-Aignan, France.
authors.forename: T
authors.initials: T
authors.lastname: Bourlard
authors.forename: M P
authors.initials: MP
authors.lastname: Bruyant-Vannier
authors.forename: A
authors.initials: A
authors.lastname: Schaumann
authors.forename: P
authors.initials: P
authors.lastname: Bruyant
authors.forename: C
authors.initials: C
authors.lastname: Morvan
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D019597:Flax / Q000201:enzymology*; D007525:Isoelectric Focusing; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D019920:Photoaffinity Labels; D053830:Plant Viral Movement Proteins; D012995:Solubility; D013379:Substrate Specificity; D014764:Viral Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D019920:Photoaffinity Labels; D053830:Plant Viral Movement Proteins; D014764:Viral Proteins