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Remarkably, two "hotspots" on TCR VB1OE (Leu32 in CDR1 and Tyr52 in CDR2, see Table 6) interact with DR4α Gln57 and Ala61 (Table 4) which form conserved contacts with TCR Vβ domain [29].
Most contacts between TCR and peptide occur through the CDR3 loops, which exhibit the greatest degree of variability whereas the preponderance of generally conserved contacts with the MHC α helices is mediated through CDR1 and CDR2 [14].
The list of contacts obtained for the model is compared to the list of 24 conserved contacts.
The basic assumption in our GLAS scoring function is that high-quality GPCR models must show a large number of these 24 conserved contacts.
Structures of TCR pMHC complexes have not shown conserved contacts that could mediate an MHC bias, although structures of different receptors sharing common variable domains suggest types of interaction that may be conserved between particular gene segment MHC pairs [ 8, 71], and, as noted above, recent results are compatible with the germline β-chain loops driving the initial pMHC contact [ 7].
The EpsE subunits form a right-handed helical arrangement in the crystal with extensive and conserved contacts between the C and N domains of neighboring subunits.
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Of note, several TCR Vβ residues make contacts to HLA-DR4 Gln57α, Ala61α and Gln70β, which stand out as MHC class II conserved contact residues from the analysis of TCR/p/MHC crystallographic complexes [29].
The highly conserved Ile-1174 contactseveralal other residues in α6 of the DH domain, including Lys-1420, Ser-1423, Asp-1424, and Leu-1426.
Conserved NTP contacts indicate a universal mechanism of NTP selection.
Two examples of such conserved predicted contacts are between Val36 resp.
Due to the speed of data collection (chemical shift information can be acquired in <5 min), low protein concentration (40 50 μM), and conserved aromatic contacts, we anticipate this method will be well-suited for bromodomain fragment screening.
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