title: Cloning, expression, purification, crystallization and preliminary crystallographic analysis of the N-terminal domain of serine glutamate repeat A (SgrA) protein from Enterococcus faecium.
abstract: Serine glutamate repeat A (SgrA) protein is an LPxTG surface adhesin of Enterococcus faecium and is the first bacterial nidogen-binding protein identified to date. It has been suggested that it binds to human nidogen, the extracellular matrix molecule of basal lamina, and plays a key role in the invasion and colonization of eukaryotic host cells. SgrA(28-288), having both a putative ligand-binding A domain and repetitive B domain, was expressed in Escherichia coli and purified using Ni-affinity and hydrophobic interaction chromatography. Further, the putative ligand-binding region, rSgrA(28-153), was subcloned, overexpressed and purified in both native and selenomethionine-derivative forms. The native rSgrA(28-153) protein crystallized in the monoclinic space group P2(1) and diffracted to 3.3 Å resolution using an in-house X-ray source, with unit-cell parameters a = 35.84, b = 56.35, c = 60.20 Å, β = 106.5°.
authors.affiliation: Centre of Advanced Study in Crystallography and Biophysics, University of Madras, Guindy Campus, Chennai 600 025, India.
authors.forename: Revathi
authors.initials: R
authors.lastname: Nagarajan
authors.forename: Antoni P A
authors.initials: AP
authors.lastname: Hendrickx
authors.forename: Karthe
authors.initials: K
authors.lastname: Ponnuraj
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D016984:Enterococcus faecium / Q000737:chemistry*; D015870:Gene Expression
chemical_list: D001426:Bacterial Proteins
keywords: Enterococcus faecium; LPxTG surface protein; extracellular matrix; surface adhesin
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