Table II.

The effect of CD1d alanine substitutions on NKT TCR binding


NKT15 Kdeq
NKT15 contact

Effect on affinity
 μM    
WT human CD1d 0.58 ± 0.13  
Arg79Aa 0.35 ± 0.02 Asp94α, Arg95α, Arg103αb Jα 
Glu83Aa >25 Tyr48β, Tyr50β 2β *** 
Lys86Aa 2.83 ± 0.13 Tyr48β 2β ** 
Met87Aa 7.9 ± 0.32 Tyr50β 2β *** 
Val147Aa 3.6 ± 0.17 Ser97α, Leu99α Jα ** 
Gln150A 2.5 ± 0.19 Gly96α, Ser97α, Thr98α, Tyr103β Jα ** 

NKT15 Kdeq
NKT15 contact

Effect on affinity
 μM    
WT human CD1d 0.58 ± 0.13  
Arg79Aa 0.35 ± 0.02 Asp94α, Arg95α, Arg103αb Jα 
Glu83Aa >25 Tyr48β, Tyr50β 2β *** 
Lys86Aa 2.83 ± 0.13 Tyr48β 2β ** 
Met87Aa 7.9 ± 0.32 Tyr50β 2β *** 
Val147Aa 3.6 ± 0.17 Ser97α, Leu99α Jα ** 
Gln150A 2.5 ± 0.19 Gly96α, Ser97α, Thr98α, Tyr103β Jα ** 

* represents CD1d alanine substitution with <4-fold reduction in binding to NKT TCR compared to the WT value; ** represents CD1d alanine substitution resulting in a 4–6-fold reduction in binding to NKT TCR compared to the WT value; and *** represents CD1d alanine substitution resulting in a >10-fold reduction in binding to NKT TCR compared to the WT value.

a

Conserved between human and mouse CD1d.

b

Denotes contacts that are not maintained in the second NKT TCR–CD1d–α-GalCer complex within the asymmetric unit.

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