DC Field | Value | Language |
dc.contributor.author | Maue, Lars | |
dc.contributor.author | Meissner, Derek | |
dc.contributor.author | Merzendorfer, Hans | |
dc.date.accessioned | 2021-12-23T16:04:27Z | - |
dc.date.available | 2021-12-23T16:04:27Z | - |
dc.date.issued | 2009 | |
dc.identifier.issn | 09651748 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/6428 | - |
dc.description.abstract | Chitin formation depends on the activity of a family II glycosyltransferase known as chitin synthase, whose biochemical and structural properties are largely unknown. Previously, we have demonstrated that the chitin portion of the peritrophic matrix in the midgut of the tobacco hornworm, Manduca sexta, is produced by chitin synthase 2 (CHS-2), one of two isoenzymes encoded by the Chs-1 and Chs-2 genes (also named Chs-A and Chs-B). and that CHS-2 is located at the apical tips of the brush border microvilli. Here we report the purification of the chitin synthase from the Manduca midgut as monitored by its activity and immuno-reactivity with antibodies to the chitin synthase. After gel permeation chromatography, the final step of the developed purification protocol, the active enzyme eluted in a fraction corresponding to a molecular mass between 440 and 670 kDa. Native PAGE revealed a single, immunoreactive band of about 520 kDa, thrice the molecular mass of the chitin synthase monomer. SDS-PAGE and immunoblotting indicated finally that an active, oligomeric complex of the chitin synthase was purified. In summary, the chitin synthase from the midgut of Manduca may prove to be a good model for investigating the enzymes' mode of action. (C) 2009 Elsevier Ltd. All rights reserved. | |
dc.description.sponsorship | Deutsche ForschungsgemeinschaftGerman Research Foundation (DFG) [SFB 431, GRK 612]; The authors are grateful to Fuensanta Martinez Rucobo and Dr. Anke Terwisscha van Scheltinga (Max-Planck-Institute for Biophysics, Frankfurt, Germany) for their help in analyzing the CHS complexes by electron microscopy. The technical help of Margret Duvel and Ulla Madler is also gratefully acknowledged. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 431 and GRK 612). | |
dc.language.iso | en | |
dc.publisher | PERGAMON-ELSEVIER SCIENCE LTD | |
dc.relation.ispartof | INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY | |
dc.subject | Biochemistry & Molecular Biology | |
dc.subject | CANDIDA-ALBICANS | |
dc.subject | Chitin | |
dc.subject | Chitin synthase | |
dc.subject | Entomology | |
dc.subject | EXPRESSION | |
dc.subject | GENES | |
dc.subject | Manduca sexta | |
dc.subject | MANDUCA-SEXTA | |
dc.subject | MEMBRANE | |
dc.subject | Midgut | |
dc.subject | Oligomerization | |
dc.subject | Peritrophic matrix | |
dc.subject | SACCHAROMYCES-CEREVISIAE | |
dc.subject | SOLUBILIZATION | |
dc.subject | SYNTHETASE | |
dc.subject | YEAST | |
dc.subject | ZYMOGENIC NATURE | |
dc.title | Purification of an active, oligomeric chitin synthase complex from the midgut of the tobacco hornworm | |
dc.type | journal article | |
dc.identifier.doi | 10.1016/j.ibmb.2009.06.005 | |
dc.identifier.isi | ISI:000269816500008 | |
dc.description.volume | 39 | |
dc.description.issue | 9 | |
dc.description.startpage | 654 | |
dc.description.endpage | 659 | |
dc.identifier.eissn | 18790240 | |
dc.publisher.place | THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND | |
dcterms.isPartOf.abbreviation | Insect Biochem. Mol. Biol. | |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | MeHa731 | - |