title: The properties and localization of Saprolegnia monoica chitin synthase differ from those of other fungi.
abstract: The presence of non-fibrillar alpha-chitin in cellulosic fungi (class Oomycetes) poses intriguing questions as to its role, subcellular localization and evolutionary significance. Previous studies reported on the similarity of chitin synthase from Saprolegnia monoica with that of other fungi. The present work describes important dissimilarities. There was no evidence that the chitin synthase of S. monoica was present in small low-density vesicles (chitosomes). Chitin synthase sedimented with membranous components of high specific gravity (sp. gr. 1.177) that could be partially but distinctly separated from membranes harbouring most of the 1,3-beta-glucan synthase in the cell (sp. gr. 1.158). In contrast to other fungi, the chitin synthase from S. monoica was greatly stimulated by digitonin: both membrane-bound and dissociated chitin synthase showed little activity in the absence of digitonin. As in other fungi, the chitin synthase from S. monoica was solubilized by digitonin and remained zymogenic after dissociation. However, unlike the enzyme from other fungi, the solubilized chitin synthase of S. monoica had a lower sedimentation coefficient, was not stimulated by phospholipids and was not inhibited by high concentrations of digitonin. Unlike the enzyme from Mucor rouxii, the solubilized chitin synthase from S. monoica did not bind to a cation exchanger. The enzyme was partially purified by four-step scheme that included sucrose density-gradient centrifugation, a single passage through a strong anion exchanger and two consecutive passages through a weak anion exchanger. The final preparation contained five to seven polypeptide bands that cochromatographed with the chitin synthase activity, some of which may be part of a presumed chitin synthase macromolecular complex.
authors.affiliation: Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato, Guanajuato, México.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Leal-Morales
authors.affiliation: Centre de Génétique Moléculaire et Cellulaire, UMR CNRS-5534 Université Lyon I, Villeurbanne, France.
authors.forename: Lucien
authors.initials: L
authors.lastname: Gay
authors.affiliation: Centre de Génétique Moléculaire et Cellulaire, UMR CNRS-5534 Université Lyon I, Villeurbanne, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Fèvre
authors.affiliation: Department of Plant Pathology, University of California, Riverside, CA 92521, USA.
authors.forename: Salomón
authors.initials: S
authors.lastname: Bartnicki-García
mesh_terms: D002687:Chitin Synthase / Q000032:analysis / Q000378:metabolism*; D005658:Fungi / Q000201:enzymology*; D013045:Species Specificity
chemical_list: D002687:Chitin Synthase