Crystallization and X-ray Crystallographic Analysis of the Adhesive SpaC Pilin Subunit in the SpaCBA Pilus of Gut-adapted Lactobacillus rhamnosus GG.
2016
Gram-positive Lactobacillus rhamnosus GG, a gut-adapted commensalic (and probiotic)
strain, is known to express sortase-assembled pili on its cell surface. These SpaCBA-called pili consist
of three different types of building blocks; the SpaA backbone-pilin subunit and the SpaB and SpaC
ancillary pilins. SpaC is a relatively large (~90kDa) multi-domain fimbrial adhesin, and while it is located
primarily at the SpaCBA pilus tip, occasionally, it can also be detected throughout the length of
pilus backbone. Functionally, SpaC mainly accounts for SpaCBA pilus-mediated interactions with intestinal
mucus, collagen, and human gut epithelial cells. Moreover, SpaC adhesiveness is also perceived
to have a causal relationship with SpaCBA pilus-induced host-cell immune responses. In order to improve the
mechanistic understanding of SpaC and its adhesive properties by structural investigation, we purified and successfully
crystallized a recombinant construct of the near full-length SpaC protein (residues 36-856) in the presence of magnesium
ions. X-ray diffraction data were collected to 2.6 A resolution. The SpaC crystal belongs to the space group P2 1 2 1 2 with
unit cell parameters a = 116.5, b = 128.3, c = 136.5 A and contains two molecules in the asymmetric unit. Presence of
conserved metal ion-dependent adhesion site containing von Willebrand factor type A domain suggests its likely role in
the function of SpaC.
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