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. 2016 Mar 11;113(12):3383–3388. doi: 10.1073/pnas.1525747113

Fig. S5.

Fig. S5.

Catalytic activity and chloroplast binding properties of mutant 13-LOX containing His/Asn-to-Ala substitutions at positions H587, H592, H777, and N781. (A) Activity measurements carried out with LeA, LA, and ARA (AR). Quantification refers to respective 13(S)-hydroperoxy products identified by HPLC and shows no detectable activity on any tested substrate. White and gray bars refer to reactions containing the full-length His/Asn-to-Ala-13-LOX (lanes 2, 4, and 6) and respective transit peptide-less 13-LOX-ΔTP mutant (lanes 1, 3, and 5) proteins. (B) Membrane-binding assays carried out with chloroplasts isolated from 7-d-old, freshly harvested barley plants. White and gray bars refer to the amount of chloroplast-bound full-length 13-LOX and 13-LOX-ΔTP that was determined for the wild-type 13-LOX (lanes 1 and 2) and the His/Asn-to-Ala-13-LOX mutant protein (lanes 3 and 4).