Open AccessExpression, purification, crystallization and preliminary crystallographic analysis of PilA from the nontypeable Haemophilus influenzae type IV pilus
Author(s) -
Kolappan Subramaniapillai,
Tracy Erin N.,
Bakaletz Lauren O.,
Munson Robert S.,
Craig Lisa
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111043910
Subject(s) - pilin , pilus , crystallization , haemophilus influenzae , microbiology and biotechnology , crystallography , protein subunit , biology , biofilm , fimbriae proteins , chemistry , virulence , bacteria , genetics , gene , antibiotics , organic chemistry
The type IV pili of nontypeable Haemophilus influenzae (NTHi) are involved in twitching motility, adherence, competence and biofilm formation. They are potential virulence factors for this important human pathogen and are thus considered to be vaccine targets. To characterize these pili, an attempt to solve the atomic structure of the major pilin subunit PilA was initiated. A 1.73 Å resolution X‐ray diffraction data set was collected from native N‐terminally truncated PilA (ΔN‐PilA). Data processing indicated a hexagonal crystal system, which was determined to belong to space group P 6 1 or P 6 5 based on the systematic absences and near‐perfect twinning of the crystal. The unit‐cell parameters were a = b = 68.08, c = 197.03 Å with four molecules in the asymmetric unit, giving a solvent content of 50%. Attempts to solve the ΔN‐PilA structure by molecular replacement with existing type IV pilin and type II secretion pseudopilin structures are in progress.