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There are no populated intermediates during the O–C conformational transition of monomeric Mad2 R133A/Q134E. Overlay of the methyl region of the 1H/13C HSQC spectra of O-Mad2 R133A/Q134E acquired at the indicated times after warming the protein to 30 °C. A series of HSQC spectra each lasting 19 min were taken for over 60 h, and only three representative spectra are shown (t = 57 min, black contours; t = 209 min, red contours; t = 1,387 min, blue contours).
