Fig. 6: One-pot synthesis of peptidyl-RNA in water.
From: Thioester-mediated RNA aminoacylation and peptidyl-RNA synthesis in water
a, Incubation of 16 with thioester 1 and thioacid 11 yields aminoacyl-RNA 17, and in situ oxidation of 11 converts 17 into peptidyl-RNA 40 in near-quantitative yield. R and R1 = peptide side chain. R2 = Me or peptide. B = nucleobase. b, 1H NMR spectra of: (1) 16A (20 mM) and 33 (80 mM) in MES buffer (500 mM, pH 6.5) after 2 h; (2) after addition of K3Fe(CN)6 (150 mM) and thioacid 11Gly (50 mM), which yields \({{\bf{40}}}_{{\bf{A}}{\bf{r}}{\bf{g}}{\bf{G}}{\bf{l}}{\bf{y}}{\bf{A}}{\bf{c}}}^{{\bf{A}}}\) (93%); (3) 16A (20 mM) and 33 (80 mM) in MES buffer (500 mM, pH 6.0) after 3 h; (4) after addition of K3Fe(CN)6 (450 mM) and Ac-Ala-Ala-SH (11AlaAla, 150 mM), which furnished \({{\bf{40}}}_{{\bf{A}}{\bf{r}}{\bf{g}}{\bf{A}}{\bf{l}}{\bf{a}}{\bf{A}}{\bf{l}}{\bf{a}}{\bf{A}}{\bf{c}}}^{{\bf{A}}}\) (89%); (5) adenosine-3′,5′-cyclic phosphate 31A (20 mM) and 33 (80 mM) in MES buffer (500 mM, pH 6.5) after 1 h, which yields aminoacyl-RNA 41; (6) after addition of K3Fe(CN)6 (300 mM) and Ac-Gly-Gly-Gly-SH (11GlyGlyGly, 100 mM), which furnished \({{\bf{42}}}_{{\bf{A}}{\bf{r}}{\bf{g}}{\bf{G}}{\bf{l}}{\bf{y}}{\bf{G}}{\bf{l}}{\bf{y}}{\bf{G}}{\bf{l}}{\bf{y}}{\bf{A}}{\bf{c}}}^{{\bf{A}}}\) (95%); see Extended Data Figs. 1 and 8 for structures of 31, 41 and 42; (7) 16A (20 mM) and l-\({{\bf{1}}}_{{\bf{L}}{\bf{e}}{\bf{u}}}^{{\bf{e}}}\) (200 mM) in MES buffer (1 M, pH 6.5) after 24 h; (8) after addition of K3Fe(CN)6 (1.05 M) and 11Gly (350 mM), which furnished peptidyl-RNA \({{\bf{40}}}_{{\bf{L}}{\bf{e}}{\bf{u}}{\bf{G}}{\bf{l}}{\bf{y}}{\bf{A}}{\bf{c}}}^{{\bf{A}}}\) (95%). 5.81 ppm (s, 1H) resonance = glycyl-thioacid by-product (see ref. 24). c, 1H NMR spectra to show: (1) 16A (20 mM); (2) 16A (20 mM), 33 (80 mM) and 11Gly (100 mM) in MES buffer (1 M, pD 6.5) after 2 h; (3) after addition of K3Fe(CN)6 (300 mM), which yields \({{\bf{40}}}_{{\bf{A}}{\bf{r}}{\bf{g}}{\bf{G}}{\bf{l}}{\bf{y}}{\bf{A}}{\bf{c}}}^{{\bf{A}}}\) (96%). For a comparable one-pot peptidyl-RNA synthesis, in which 16A, 33 and K3Fe(CN)6 are incubated to yield 17 and then thioacid 11Gly was added at 2 h to furnish 40, see Supplementary Fig. 250.
