Modeling of NAR V and comparison with camel and
human VH structures. (a) The
VH/VL of the KOL human crystal (18, 23)
showing the disulfide bridge in the CDR3 VH (boxed). Note
that the VL (left) binding site is obscured by the
VH CDR3. Canonical intradomain disulfide bonds between the
two β sheets of the H and L chains are displayed in yellow (not
boxed). The boxed disulfide bridge (also yellow) is postulated to be
similar to bonds that would form in NAR Type I CDR3. (b)
Crystal structure of the unusual camel VH (15). The
disulfide bridge between CDR1 and 3 is boxed, and the canonical
intradomain disulfide bridge is displayed. The disulfide bond displayed
here is postulated to be similar to those formed by CDR1 and 3 Cys
residues in NAR Type II proteins. (c and
d) Model of the Type I NAR V domain (red) superimposed
on the camel VH (white). Note the high similarity of the
two structures except in the CDR2 region, which in NAR is very small
and may connect the two β sheets in a fashion similar to Ig C1
domains (33). The canonical disulfide bridge (yellow) and serine (red)
in NAR CDR1 (S-28, Fig. 2) are displayed for bearings.
(e and f) Potential disulfide
bond formed in NAR between unconventional Cys (boxed) in FR2 (C-35) and
FR 4 (C-107); see Figs. 2 and 3
a and c.
The side chains of these two Cys project into the region in which
conventional VH and VL interact. We propose
that the small size of the highly conserved glycine at position 84
(boxed G-84, Figs. 2 and 3c) permits C-35 and C-107 to
disulfide-bond; it is important to emphasize that this glycine is
conserved only in Type I NAR sequences and is rarely found in Ig/TCR
of other vertebrates or even in Type II NAR (3). We speculate that NAR
is more compact, and, because of irregular H bonding in the G strand
(FR4) of the model, it is possible that a disulfide bond can be formed
between C-35 and C-107. The canonical disulfide bond (C-22, C-83) and
the invariant tryptophan (W-36) forming the core of the NAR domain, as
well as the invariant Gly-Xaa-Gly (ref. 18; G-108, G-110), are
displayed.