Sucrose synthase is an integral component of the cellulose synthesis machinery.
Cellulose synthesis in plants is believed to be carried out by the plasma membrane-associated rosette structure which can be observed by electron microscopy. Despite decade-long speculation, it had not been demonstrated whether the rosette is the site of catalytic activity of cellulose synthesis. To determine the relationship between this structure and cellulose synthesis, we successfully isolated detergent-insoluble rosettes from the plasma membrane of bean epicotyls. However, the purified rosettes did not possess cellulose synthesis activity in vitro. Conversely, detergent-soluble granular particles of approximately 9.5-10 nm diameter were also isolated and exhibited UDP-glucose binding activity and possessed beta-1,4-glucan (cellulose) synthesis activity in vitro. The particle, referred to as the catalytic unit of cellulose synthesis, was enriched with a 78 kDa polypeptide which was verified as sucrose synthase like by mass spectrometry and immunoblotting. The catalytic units were able to bind to the rosettes and retained the cellulose synthesis activity in the presence of UDP-glucose or sucrose plus UDP when supplemented with magnesium. The incorporation of the catalytic unit into the rosette structure was confirmed by immunogold labeling with anti-sucrose synthase antibodies under an electron microscope. Our results suggest that the plasma membrane-associated rosette anchors the catalytic unit of cellulose synthesis to form the functional cellulose synthesis machinery.
Fujii
Satoshi
S
Department of Biological Sciences, Graduate School of Science, Osaka University, Osaka, 560-0043 Japan. f ujii@bio.sci.osaka-u.ac.jp
Hayashi
Takahisa
T
Mizuno
Koichi
K
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism; D002482:Cellulose / Q000096:biosynthesis*; D007887:Fabaceae / Q000201:enzymology*; D005936:Glucans / Q000096:biosynthesis; D005964:Glucosyltransferases / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013395:Sucrose / Q000378:metabolism; D014532:Uridine Diphosphate Glucose / Q000378:metabolism
D005936:Glucans; D013395:Sucrose; D002482:Cellulose; C040088:1,4-glucan; D005964:Glucosyltransferases; C022699:sucrose synthase; D014532:Uridine Diphosphate Glucose