Extended Data Fig. 3: Characterization of the Tri22-catalyzed reaction product, 11.

a) Schematic of a biochemical assay performed containing HAA, succinyl-CoA, Tri29-31, and Tri22 that is subjected to base hydrolysis, followed by OPTA derivatization to yield 2-HYAA-D. b) EICs showing the generation of 2-HYAA-D from the biochemical assay previously described in comparison with a derivatized 2-HYAA synthesized standard. Omission of Tri22, HAA, or succinyl-CoA from the couple enzymatic reaction resulted in abolishment of 2-HYAA-D. The calculated mass for 2-HYAA-D: m/z = 265.0642 ([M + H]⁺) is used for each trace. A 10-ppm mass error tolerance was used for each trace. At least three independent replicates were performed for each assay, and representative results are shown.