Crystallization and X-ray diffraction analysis of a novel surface-adhesin protein: protein E from Haemophilus influenzae.

Singh, BirendraLund University,Lunds universitet,Klinisk mikrobiologi, Malmö,Forskargrupper vid Lunds universitet,Clinical Microbiology, Malmö,Lund University Research Groups (författare)

Protein E (PE) is a ubiquitous multifunctional surface protein of Haemophilus spp. and other bacterial pathogens of the Pasteurellaceae family. H. influenzae utilizes PE for attachment to respiratory epithelial cells. In addition, PE interacts directly with plasminogen and the extracellular matrix (ECM) components vitronectin and laminin. Vitronectin is a complement regulator that inhibits the formation of the membrane-attack complex (MAC). PE-mediated vitronectin recruitment at the H. influenzae surface thus inhibits MAC and protects against serum bactericidal activity. Laminin is an abundant ECM protein and is present in the basement membrane that helps in adherence of H. influenzae during colonization. Here, the expression, purification and crystallization of and the collection of high-resolution data for this important H. influenzae adhesin are reported. To solve the phase problem for PE, Met residues were introduced and an SeMet variant was expressed and crystallized. Both native and SeMet-containing PE gave plate-like crystals in space group P2(1), with unit-cell parameters a = 44, b = 57, c = 61 Å, β = 96°. Diffraction data collected from native and SeMet-derivative crystals extended to resolutions of 1.8 and 2.6 Å, respectively.

Tamim, Al-JubairLund University,Lunds universitet,Klinisk mikrobiologi, Malmö,Forskargrupper vid Lunds universitet,Clinical Microbiology, Malmö,Lund University Research Groups(Swepub:lu)med-tmj (författare)
Förnvik, KarolinaLund University,Lunds universitet,Geriatrik,Forskargrupper vid Lunds universitet,Geriatric Medicine,Lund University Research Groups,Geriatrics(Swepub:lu)mbfy-kaf (författare)
Thunnissen, Marjolein (författare)
Riesbeck, KristianLund University,Lunds universitet,Klinisk mikrobiologi, Malmö,Forskargrupper vid Lunds universitet,Clinical Microbiology, Malmö,Lund University Research Groups(Swepub:lu)mikr-kri (författare)
Klinisk mikrobiologi, MalmöForskargrupper vid Lunds universitet (creator_code:org_t)

Ingår i:Acta Crystallographica. Section F: Structural Biology and Crystallization Communications68:Pt 2, s. 222-2262053-230X

Av författaren/redakt...: Singh, Birendra; Tamim, Al-Jubair; Förnvik, Karolin ...; Thunnissen, Marj ...; Riesbeck, Kristi ...

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