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For evaporation of this water to occur, the hydrogen bonds between the individual water molecules must be broken, requiring a relatively high amount of energy that in part includes heat

A key feature of the inactive state is the presence of a major hydrogen bond network (MHN) involving residues N111 3.35 , N295 7.46 , D74 2.50 and N298 7.49 (superscripts denote BallesterosWeinstein numbering 96 ), which acts as a molecular switch that stabilizes the inactive conformation

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coli induced with 0.5 mM IPTG and grown overnight at 20C at 150 rpm