In vitro synthesis of cellulose microfibrils by a membrane protein from protoplasts of the non-vascular plant Physcomitrella patens

2015 ◽  
Vol 470 (2) ◽  
pp. 195-205 ◽  
Author(s):  
Sung Hyun Cho ◽  
Juan Du ◽  
Ian Sines ◽  
Venkata Giridhar Poosarla ◽  
Venkata Vepachedu ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Physcomitrella Patens ◽  
Vascular Plant ◽  
Cellulose Synthase ◽  
Cellulose Microfibrils ◽  
Protein Preparation ◽  
In Vitro Synthesis

A membrane protein preparation isolated from moss protoplasts overexpressing the moss cellulose synthase 5 (CesA5), produced cellulose microfibrils. The microfibrils were frequently attached to putative CesA, suggesting that the moss is a useful system for studying plant CesA.

scholarly journals In vitro synthesis of the major lens membrane protein.

1980 ◽  
Vol 77 (2) ◽  
pp. 725-729 ◽  
Author(s):  
F. C. Ramaekers ◽  
A. M. Selten-Versteegen ◽  
E. L. Benedetti ◽  
I. Dunia ◽  
H. Bloemendal
Keyword(s):  
Membrane Protein ◽  
In Vitro Synthesis

IN VITRO SYNTHESIS OF CELLULOSE MICROFIBRILS BY DETERGENT EXTRACTS OF PLANT MICROSOMAL FRACTIONS

2002 ◽  
Author(s):  
Josephine Lai Kee Him ◽  
Henri Chanzy ◽  
Martin Muller ◽  
Jean-Luc Putaux ◽  
Vincent Bulone
Keyword(s):  
Cellulose Microfibrils ◽  
In Vitro Synthesis

scholarly journals A single heterologously expressed plant cellulose synthase isoform is sufficient for cellulose microfibril formation in vitro

2016 ◽  
Vol 113 (40) ◽  
pp. 11360-11365 ◽  
Author(s):  
Pallinti Purushotham ◽  
Sung Hyun Cho ◽  
Sara M. Díaz-Moreno ◽  
Manish Kumar ◽  
B. Tracy Nixon ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Wall ◽  
Secondary Cell Wall ◽  
Cellulose Synthase ◽  
Fiber Formation ◽  
Cellulose Microfibrils ◽  
Cell Wall Formation ◽  
Secondary Cell Wall Formation ◽  
Wall Formation

Plant cell walls are a composite material of polysaccharides, proteins, and other noncarbohydrate polymers. In the majority of plant tissues, the most abundant polysaccharide is cellulose, a linear polymer of glucose molecules. As the load-bearing component of the cell wall, individual cellulose chains are frequently bundled into micro and macrofibrils and are wrapped around the cell. Cellulose is synthesized by membrane-integrated and processive glycosyltransferases that polymerize UDP-activated glucose and secrete the nascent polymer through a channel formed by their own transmembrane regions. Plants express several different cellulose synthase isoforms during primary and secondary cell wall formation; however, so far, none has been functionally reconstituted in vitro for detailed biochemical analyses. Here we report the heterologous expression, purification, and functional reconstitution of Populus tremula x tremuloides CesA8 (PttCesA8), implicated in secondary cell wall formation. The recombinant enzyme polymerizes UDP-activated glucose to cellulose, as determined by enzyme degradation, permethylation glycosyl linkage analysis, electron microscopy, and mutagenesis studies. Catalytic activity is dependent on the presence of a lipid bilayer environment and divalent manganese cations. Further, electron microscopy analyses reveal that PttCesA8 produces cellulose fibers several micrometers long that occasionally are capped by globular particles, likely representing PttCesA8 complexes. Deletion of the enzyme’s N-terminal RING-finger domain almost completely abolishes fiber formation but not cellulose biosynthetic activity. Our results demonstrate that reconstituted PttCesA8 is not only sufficient for cellulose biosynthesis in vitro but also suffices to bundle individual glucan chains into cellulose microfibrils.

In vitro synthesis and assembly of a 68 kDa outer mitochondrial membrane protein from rat liver

1985 ◽  
Vol 814 (1) ◽  
pp. 35-42 ◽  
Author(s):  
Carole Noël ◽  
Vasilios Nicolaou ◽  
Carole Argan ◽  
Richard A. Rachubinski ◽  
Gordon C. Shore
Keyword(s):  
Membrane Protein ◽  
Rat Liver ◽  
Mitochondrial Membrane ◽  
Outer Mitochondrial Membrane ◽  
In Vitro Synthesis ◽  
Mitochondrial Membrane Protein
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