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The human NKT TCR was more tolerant to modifications at the 4′-OH position and less tolerant to modifications in the 3′-OH position of α-GalCer. Within the human NKT TCR-CD1d-α-GalCer complex, the 4′-OH group hydrogen-bonds to the main chain of Phe-29α, whereas the 3′-OH hydrogen-bonds to Ser-30α. The human NKT TCR-CD1d-4′,4″-deoxy-α-GalCer complex showed that there were limited perturbations arising from the loss of the interaction between the 4′-OH moiety and Phe-29.
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