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Fig. 3 In vitro reconstitution and kinetic analysis of Q-glycosylation (A) In vitro reconstitution of galQ34 in human tRNATyr with recombinant QTGAL in the presence (+) or absence (−) of UDP-gal. (B) In vitro reconstitution of manQ34 in E. coli tRNAAsp with recombinant QTMAN in the presence (+) or absence (−) of GDP-man. (C and D) Kinetic analyses of galQ34 (C) or manQ34 (D) formation by recombinant QTGAL or QTMAN with different concentrations of human tRNATyr or E. coli tRNAAsp (upper) and UDP-gal or GDP-man (lower), respectively. The initial velocity of galQ34 or manQ34 formation (nM/s) is plotted as the mean ± standard deviation (SD; n = 3). Data are fitted to the Michaelis-Menten equation to yield Km and Vmax values (inset). See also Figures S1, S10, and S11.
Reprinted from Cell, 186(25), Zhao, X., Ma, D., Ishiguro, K., Saito, H., Akichika, S., Matsuzawa, I., Mito, M., Irie, T., Ishibashi, K., Wakabayashi, K., Sakaguchi, Y., Yokoyama, T., Mishima, Y., Shirouzu, M., Iwasaki, S., Suzuki, T., Suzuki, T., Glycosylated queuosines in tRNAs optimize translational rate and post-embryonic growth, 5517-5535.e24, Copyright (2023) with permission from Elsevier. Full text @ Cell