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Once the stopper loses vacuum seal (no audible hiss on first puncture), air exchange is occurring refrigeration alone cannot compensate for oxidation at that stage.
The site includes Glu170 as well as candidate residues Gln143 (r 2 ) in helix 3, Arg152 (r 7 ) and Thr154-Pro156 in a loop preceding strand 3, and Gly167-Thr169 (r 46 ) in helix 4 (Figs 1c and 6a)
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