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. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: Phytochemistry. 2014 Nov 4;112:151–159. doi: 10.1016/j.phytochem.2014.09.015

Figure 5. In vitro crosslinking assays using tomato P1 extensin.

Figure 5

Extensin crosslinking assays (30min) were performed using tomato P1 extensin substrate to test the activities of folded CG5-6xHis. Control assays included folding solution (A), water (B), corresponding proteins isolated from E. coli transformed with the empty vector-EV (C), and horseradish peroxidase (D). Extensin polymerization was observed when either CG5-6xHis (E) or the tomato pI 4.6 EP (F) was added. Crosslinking is observed by the size shift in P1 monomer by gel permeation chromatography upon forming P1 oligomer and P1 polymer with concomitant decrease in P1 monomer. Extended incubation times leave much of the P1 polymer formed in (E) and (F) insoluble (e.g. not detected by gel-filtration).