Crystallization and X-ray crystallographic analysis of the adhesive SpaC pilin subunit in the SpaCBA pilus of gut-adapted Lactobacillus rhamnosus GG

Abhiruchi Kant, Ingemar von Ossowski, Airi Palva, Vengadesan Krishnan*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

Abstract

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 (similar to 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 magnesiumions. X-ray diffraction data were collected to 2.6 resolution. The SpaC crystal belongs to the space group P21212 with unit cell parameters a = 116.5, b = 128.3, c = 136.5 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.

Original languageEnglish
Pages (from-to)365-371
Number of pages7
JournalPROTEIN AND PEPTIDE LETTERS
Volume23
Issue number4
DOIs
Publication statusPublished - 2016
MoE publication typeA1 Journal article-refereed

Keywords

  • Adhesin
  • Lactobacillus rhamnosus GG
  • Pili
  • Fimbria
  • Probiotics
  • SpaC
  • GRAM-POSITIVE BACTERIA
  • FUNCTIONAL-ANALYSIS
  • STRUCTURAL BASIS
  • PYOGENES PILUS
  • INTEGRIN
  • COLLAGEN
  • BINDING
  • MODEL
  • PURIFICATION
  • RECOGNITION

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