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Fig. 5 | Plant Methods

Fig. 5

From: A plant protein farnesylation system in prokaryotic cells reveals Arabidopsis AtJ3 produced and farnesylated in E. coli maintains its function of protecting proteins from heat inactivation

Fig. 5

OsDjA4 was farnesylated in OsPFT-producing E. coli cells. a OsDjA4 produced in E. coli cells harboring either pGEX-OsDjA4 alone (OsDjA4) or both pGEX-OsDjA4 and pRSF-Duet1-OsPFT (OsDjA4 + OsPFT) was purified using Glutathione Sepharose 4B beads and run in duplicate, one for Coomassie blue staining and one for western blot detection, using SDS-PAGE. The OsDjA4 purified from pGEX-OsDjA4 and pRSF-Duet1-OsPFT co-transformed E. coli cells exhibited faster electrophoretic mobility than that from cells harboring pGEX-OsDjA4 alone, indicating OsDjA4 from co-transformed cells was farnesylated. b The extracted ion chromatograms (EIC) of the C-terminal peptide of OsDjA4 with and without farnesylation. The target peptide (DEDDDEDAGAGPRVQC) produced from the cells containing pGEX-OsDjA4 alone (upper panel, OsDjA4) showed only a non-farnesylated form with 874.8373 (2+) m/z at 18.72 min, but the target peptide produced from the cells containing pRSF-Duet1-OsPFT and pGEX-OsDjA4 (lower panel, OsPFT + OsDjA4) showed both non-farnesylated form with 874.8373 (2+) m/z at 19.77 min and farnesylated form with 948.4208 (2+) m/z at 66.20 min, respectively

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Plant Methods

ISSN: 1746-4811