title: The mannose-specific lectin domains of Flo1p from Saccharomyces cerevisiae and Lg-Flo1p from S. pastorianus: crystallization and preliminary X-ray diffraction analysis of the adhesin-carbohydrate complexes.
abstract: Flo1p and Lg-Flo1p are two cell-wall adhesins belonging to the Flo (flocculation) protein family from the yeasts Saccharomyces cerevisiae and S. pastorianus. The main function of these modular proteins endowed with calcium-dependent lectin activity is to mediate cell-cell adhesion events during yeast flocculation, a process which is well known at the cellular level but still not fully characterized from a molecular perspective. Recently, structural features of the N-terminal Flo lectin domains, including the N-terminal domain of Lg-Flo1p (N-Lg-Flo1p), and their interactions with carbohydrate molecules have been investigated. However, structural data concerning the N-terminal domain of Flo1p (N-Flo1p), which is the most specific among the Flo proteins, are missing and information about the N-Lg-Flo1p-carbohydrate interaction still lacks detailed structural insight. Here, the crystallization and preliminary X-ray characterization of the apo form and the mannose complex of N-Flo1p and X-ray analysis of N-Lg-Flo1p crystals soaked in α-1,2-mannobiose are reported. The N-Flo1p crystals diffracted to a resolution of 1.43 Å in the case of the apo form and to 2.12 Å resolution for the mannose complex. Both crystals were orthorhombic and belonged to space group P212121, with one molecule in the asymmetric unit. The N-Lg-Flo1p-α-1,2-mannobiose complex crystal diffracted to 1.73 Å resolution and belonged to the monoclinic space group P1211 with two molecules in the asymmetric unit.
authors.affiliation: Structural Biology Brussels (SBB) - Department of Bioengineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium. fielasi@vub.ac.be
authors.forename: Francesco S
authors.initials: FS
authors.lastname: Ielasi
authors.forename: Parveen
authors.initials: P
authors.lastname: Goyal
authors.forename: Mike
authors.initials: M
authors.lastname: Sleutel
authors.forename: Alexandre
authors.initials: A
authors.lastname: Wohlkonig
authors.forename: Ronnie G
authors.initials: RG
authors.lastname: Willaert
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D001665:Binding Sites; D003460:Crystallization; D018360:Crystallography, X-Ray; D005427:Flocculation; D008351:Mannans / Q000378:metabolism*; D037241:Mannose-Binding Lectins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; C087578:FLO1 protein, S cerevisiae; D008351:Mannans; D037241:Mannose-Binding Lectins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C014647:mannobiose
keywords: Flo proteins; Flo1p; Lg-Flo1p; Saccharomyces cerevisiae; Saccharomyces pastorianus; flocculins; lectins; yeast flocculation