Kumari Yadav, Rajnesh and Krishnan, Vengadesan (2020) The adhesive PitA pilus protein from the early dental plaque colonizer \it Streptococcus oralis: expression, purification, crystallization and X-ray diffraction analysis. Acta Crystallographica Section F, 76 (1). pp. 8-13.

Text The adhesive PitA pilus protein from the early dental plaque colonizer Streptococcus oralis expression, purification, crystallization and X-ray diffraction analysis.bib - Published Version Restricted to Repository staff only Download (1914b) | Request a copy

Abstract

PitA is the putative tip adhesin of the pilus islet 2 (PI-2)-encoded sortase-dependent pilus in the Gram-positive \it Streptococcus oralis, an opportunistic pathogen that often flourishes within the diseased human oral cavity. Early colonization by \it S. oralis and its interaction with \it Actinomyces oris seeds the development of oral biofilm or dental plaque. Here, the PI-2 pilus plays a vital role in mediating adherence to host surfaces and other bacteria. A recombinant form of the PitA adhesin has now been produced and crystallized. Owing to the large size (∼100kDa), flexibility and complicated folding of PitA, obtaining diffraction-quality crystals has been a challenge. However, by the use of limited proteolysis with α-chymotrypsin, the diffraction quality of the PitA crystals was considerably enhanced to 2.16Å resolution. These crystals belonged to space group \it P1, with unit-cell parameters \it a = 61.48, \it b = 70.87, \it c~=~82.46Å, α = 80.08, β~=~87.02, γ = 87.70\circ. The anomalous signal from the terbium derivative of α-chymotrypsin-treated PitA crystals prepared with terbium crystallophore (Tb-Xo4) was sufficient to obtain an interpretable electron-density map via terbium SAD phasing.

Item Type:	Article
Uncontrolled Keywords:	adhesins, PitA, PI-2 sortase-dependent pilus, Streptococcus oralis, proteolysis, terbium crystallophore
Subjects:	Biochemical and Biophysical Sciences
Depositing User:	RCB Library
Date Deposited:	29 Jul 2020 06:34
Last Modified:	29 Jul 2020 06:34
URI:	http://rcb.sciencecentral.in/id/eprint/429

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