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. Author manuscript; available in PMC: 2020 Jan 15.
Published in final edited form as: Nat Chem Biol. 2019 Jul 15;15(8):795–802. doi: 10.1038/s41589-019-0314-6

Figure 5.

Figure 5.

In vitro reconstitution of the PKS and the glycosyltransferase proteins involved in pactamycin biosynthesis. a. Enzymatic reactions from 3ABA to GlcNac-3ABA-PtmI and to GlcNAc-β-ketoacyl-PtmI; b. Deconvoluted mass spectrum of a reaction mixture containing PtmS, PtmI, and PtmJ; c. Deconvoluted mass spectrum of a reaction mixture with PtmS, PtmI, PtmK, and PtmJ; d. Deconvoluted mass spectrum of a reaction mixture with PtmS, PtmI, PtmK, and PtmJ using 3ABA-2,4,5,6-d4 as substrate; e. Deconvoluted mass spectrum of a reaction mixture with PtmS, PtmI, PtmK, and boiled PtmJ. Magenta circle, holo-PtmI; blue circle, acetyl-PtmI; black circle, malonyl-PtmI; red diamond, 3ABA-PtmI; purple triangle, 3AP-3OP-PtmI; orange star, GlcNAc-3ABA-PtmI; yellow star, GlcNAc-3AP-3OP-PtmI. The gluconoylated proteins are labeled with “Gluc”. Deuterated species are labeled with d4. Experiments 5b-e were performed independently at least three times with similar results.