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. 2022 Jan 18;119(4):e2103099119. doi: 10.1073/pnas.2103099119

Fig. 2.

Fig. 2.

Analysis of the purity and bioactivity of multisite lipidated biopolymers. (A) Schematic representation of ELP-GFP reporter constructs with 1, 5, or 10 pAzF residues. Target positions for pAzF are indicated in red. (B) Relative abundance of detected nonstandard amino acids at target residues of ELP units based on quantitative MS. Treatment with ISAz selectively converts reduced pAzF residues, pAF, back into pAzF. (C) Click-chemistry with FA alkynes functionalizes all pAzF, but not pAF, residues (n = 3, error bars: mean ± SD). (D–G) Intact MS of full-length ELP(FA)-GFP after click-chemistry with (blue) or without (black) ISAz treatment. (H) Activity of recombinant trx, trx-ELP(10pAzF), and trx-ELP(10FA) at protein quantities ranging from 25 to 150 pmol per well. (I) Effect of HSA on the activity of recombinant trx, trx-ELP(10pAzF) and trx-ELP(10FA) using 100 pmol of each protein and 500 pmol HSA. Data are normalized to the activity of each protein without HSA (n = 3, error bars: mean ± SEM). *P < 0.05, **P < 0.01.