Related Experiment Video
Updated: Jul 16, 2026
11:08
Chemo-enzymatic Synthesis of N-glycans for Array Development and HIV Antibody Profiling
Published on: February 5, 2018
8.7K
New insights on β-glycan synthases using in vitro GT-array (i-GT-ray) platform
Matrika Bhattarai1, Qi Wang2, Zawar Hussain3
1From the Department of Environmental and Plant Biology, Ohio University, Athens, OH, 45701, USA; Molecular and Cellular Biology Program, Ohio University, Athens, OH, 45701, USA.
Plant Physiology and Biochemistry : PPB
|August 20, 2024
Summary
The novel in vitro GT-array (i-GTray) platform successfully produced soluble and active β-glycan synthases, revealing new insights into plant cell wall polysaccharide synthesis and enzyme interactions.
Area of Science:
- Plant Biology
- Biochemistry
- Molecular Biology
Background:
- Cellulose and hemicellulose are crucial structural polysaccharides in plant cell walls, synthesized by membrane-bound glycosyltransferases (GTs) known as synthases.
- These membrane proteins are challenging to purify and study in vitro, hindering research into their function.
Purpose of the Study:
- To evaluate the efficacy of the in vitro GT-array (i-GTray) platform for producing and testing β-glycan synthases.
- To investigate the biochemical properties and interactions of plant β-glycan synthases.
Main Methods:
- Developed and utilized the in vitro GT-array (i-GTray) platform for cell-free production of membrane-bound GTs.
- Tested five plant β-glycan synthases involved in cellulose, xyloglucan, and (gluco)mannan synthesis.
- Analyzed enzyme activity, product formation, and protein-protein interactions.
Main Results:
- All tested β-glycan synthases were produced in soluble, active forms using the i-GTray platform, active without detergents or membrane lipids, and did not require a primer.
- Synthases produced ethanol-insoluble products degraded by specific hydrolases.
- Direct interaction between AtCslC4 and AtXXT1 formed an active xyloglucan synthase producing xylosylated cello-oligosaccharides.
Conclusions:
- The i-GTray platform is a powerful tool for functional genomics, enabling the study of difficult-to-purify membrane proteins like β-glycan synthases.
- The study uncovered unexpected properties of β-glycan synthases and demonstrated their potential for direct interaction and complex product synthesis.
Related Concept Videos
01:27Glucose Transporters
Glucose transporters facilitate the transport of glucose across the cell membrane. In addition to glucose, some glucose transporters can also aid the movement of other hexoses such as fructose, mannose, and galactose.
Facilitated diffusion-glucose transporters (GLUTs) are encoded by the solute-linked carrier (SLC) family 2, subfamily A gene family, or SLC2A. The 14 GLUT protein members are distributed into three classes:
Facilitated diffusion-glucose transporters (GLUTs) are encoded by the solute-linked carrier (SLC) family 2, subfamily A gene family, or SLC2A. The 14 GLUT protein members are distributed into three classes:
01:26Glucose Absorption Into the Small Intestine
Complex carbohydrates consumed cannot be absorbed into the small intestine in their original form. First, they must be hydrolyzed to a monosaccharide form such as glucose or galactose. These monosaccharides are then transported across the intestinal membrane and into the blood via transcellular transport. The intestinal epithelial cells allow the movement of these monosaccharides with a defined 'entry' through membrane transporter proteins present on their apical membrane and 'exit' via the...