Upon sample injection, the mobile phase was held at 100% (v/v) solvent A for 7.14 min, then a gradient was applied with sequential composition targets of 90% (v/v) solvent A and 10% (v/v) solvent B for 21.42 min, 80% (v/v) solvent A and 20% (v/v) solvent B for 42.85 min, 20% (v/v) solvent A and 80% (v/v) solvent B for 57.14 min, 100% (v/v) solvent B for 64.28, and 100% (v/v) solvent A for 78.57 min
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This reaction produces one molecule of water (H 2 O), as well as two amino acids joined by a peptide bond (CONH)
Acknowledgements We thank Didier Vertommen for generously providing mass spectrometry services, Nicolas Foloppe, Guido Verniest, and Brandn Prez for discussion, Remy Loris and Inge Van Molle for collecting crystal diffraction data, Martine De Cock for help in preparing the manuscript, and the Proxima1 beamline scientists at SOLEIL for their technical support