Recombinant Arabidopsis thaliana 18 kDa cell wall protein
Description
Functional Properties
-
In vitro activity: Inhibits tubulin polymerization, reducing microtubule density .
-
Genetic modulation: Overexpression causes left-handed root skewing and elongated cells, while RNAi silencing shortens cells and reduces lobe formation .
Recombinant Production and Applications
While MAP18 itself is not described as a cell wall protein, studies on recombinant Arabidopsis proteins provide insights into production methodologies:
Expression Systems
-
Escherichia coli: MAP18 was expressed as a GST fusion protein in E. coli for biochemical assays .
-
Plant cell systems: Electroporation-mediated protein delivery into Arabidopsis cells with intact cell walls has been optimized for recombinant proteins .
Case Study: Recombinant MAP18
| Parameter | Details |
|---|---|
| Expression host | E. coli (pGEX-4T vector) |
| Purification | Affinity chromatography via GST tag |
| Activity | Reduces microtubule polymerization by 40% at 10 µM |
Overexpression vs. RNAi Silencing
Technical Advances in Recombinant Protein Production
Recent innovations for Arabidopsis-based systems include:
-
Super-expression platform: Yields up to 0.4 mg protein/g fresh weight, demonstrated for membrane complexes like oligosaccharyltransferase .
-
Chaperone co-expression: Enhances solubility of challenging targets like cell wall glycosyltransferases .
Challenges and Gaps
-
Nomenclature clarity: The term "18 kDa cell wall protein" may conflate MAP18 with unrelated cell wall enzymes (e.g., RGPs or ABCG18) .
-
Functional data: No direct evidence links MAP18 to cell wall biosynthesis, though its role in microtubule dynamics indirectly affects cell wall patterning .
