RRC ID 37558
Author Cho SH, Du J, Sines I, Poosarla VG, Vepachedu V, Kafle K, Park YB, Kim SH, Kumar M, Nixon BT.
Title In vitro synthesis of cellulose microfibrils by a membrane protein from protoplasts of the non-vascular plant Physcomitrella patens.
Journal Biochem. J.
Abstract Plant cellulose synthases (CesAs) form a family of membrane proteins that are associated with hexagonal structures in the plasma membrane called CesA complexes (CSCs). It has been difficult to purify plant CesA proteins for biochemical and structural studies. We describe CesA activity in a membrane protein preparation isolated from protoplasts of Physcomitrella patens overexpressing haemagglutinin (HA)-tagged PpCesA5. Incubating the membrane preparation with UDP-glucose predominantly produced cellulose. Negative-stain EM revealed microfibrils. Cellulase bound to and degraded these microfibrils. Vibrational sum frequency generation (SFG) spectroscopic analysis detected the presence of crystalline cellulose in the microfibrils. Putative CesA proteins were frequently observed attached to the microfibril ends. Combined cross-linking and gradient centrifugation showed bundles of cellulose microfibrils with larger particle aggregates, possibly CSCs. These results suggest that P. patens is a useful model system for biochemical and structural characterization of plant CSCs and their components.
Volume 470(2)
Pages 195-205
Published 2015-9-1
DOI 10.1042/BJ20141391
PII BJ20141391
PMID 26348908
MeSH Bryopsida / chemistry* Cellulose / chemistry* Glucosyltransferases / chemistry* Membrane Proteins / chemistry* Microfibrils / chemistry* Plant Lectins / chemistry Plant Proteins / chemistry* Protoplasts / chemistry*
IF 3.857
Times Cited 3
Arabidopsis / Cultured plant cells, genes pdp24095