Figure S1. Synthesis of GxTX-594. (A)... | Rockefeller University Press

Figure S1.

$Synthesis of GxTX-594.(A) Molecular model of GxTX-594. Scale bar, 10 Å. Backbone of GxTX peptide depicted with ribbon. Cys13(maleimide-Alexa 594) depicted with CPK coloring. (B) HPLC chromatogram of Ser13Cys GxTX. Gradient described in –GxTX synthesis. Ser13Cys GxTX eluted at 12.6 min, peak 1, which corresponds to 33% ACN. MALDI-TOF mass spectrometry profile of peak 1 (inset). (C) HPLC chromatogram of GxTX-594 conjugation reaction between Alexa Fluor 594-maleimide and Ser13Cys GxTX. Peak 1 is Ser13Cys GxTX (retention time, 12.8 min; 33% ACN); peak 2 is a minor product from conjugation; and peak 3 is GxTX-594, the major product from conjugation (retention time, 16.4 min; 35% ACN). The fractions corresponding to peak 3 were combined. (D) HPLC chromatogram of the combined peak 3 fractions from C, a GxTX-594 preparation used in this study. 2 μl of 13.1 μM GxTX-594 diluted in 200 μl of 0.1% TFA was injected. MALDI-TOF mass spectrometry profile of the combined peak 3 fractions (inset). a.m.u., atomic mass unit; Rel. Int., relative intensity.$

Synthesis of GxTX-594. (A) Molecular model of GxTX-594. Scale bar, 10 Å. Backbone of GxTX peptide depicted with ribbon. Cys13(maleimide-Alexa 594) depicted with CPK coloring. (B) HPLC chromatogram of Ser13Cys GxTX. Gradient described in –GxTX synthesis. Ser13Cys GxTX eluted at 12.6 min, peak 1, which corresponds to 33% ACN. MALDI-TOF mass spectrometry profile of peak 1 (inset). (C) HPLC chromatogram of GxTX-594 conjugation reaction between Alexa Fluor 594-maleimide and Ser13Cys GxTX. Peak 1 is Ser13Cys GxTX (retention time, 12.8 min; 33% ACN); peak 2 is a minor product from conjugation; and peak 3 is GxTX-594, the major product from conjugation (retention time, 16.4 min; 35% ACN). The fractions corresponding to peak 3 were combined. (D) HPLC chromatogram of the combined peak 3 fractions from C, a GxTX-594 preparation used in this study. 2 μl of 13.1 μM GxTX-594 diluted in 200 μl of 0.1% TFA was injected. MALDI-TOF mass spectrometry profile of the combined peak 3 fractions (inset). a.m.u., atomic mass unit; Rel. Int., relative intensity.