title: Crystallization and preliminary structure determination of the transfer protein TraM from the Gram-positive conjugative plasmid pIP501.
abstract: The major means of horizontal gene spread (e.g. of antibiotic resistance) is conjugative plasmid transfer. It presents a serious threat especially for hospitalized and immuno-suppressed patients, as it can lead to the accelerated spread of bacteria with multiple antibiotic resistances. Detailed information about the process is available only for bacteria of Gram-negative (G-) origin and little is known about the corresponding mechanisms in Gram-positive (G+) bacteria. Here we present the purification, biophysical characterization, crystallization and preliminary structure determination of the TraM C-terminal domain (TraMΔ, comprising residues 190-322 of the full-length protein), a putative transfer protein from the G+ conjugative model plasmid pIP501. The crystals diffracted to 2.5 Å resolution and belonged to space group P1, with unit-cell parameters a = 39.21, b = 54.98, c = 93.47 Å, α = 89.91, β = 86.44, γ = 78.63° and six molecules per asymmetric unit. The preliminary structure was solved by selenomethionine single-wavelength anomalous diffraction.
authors: Institute for Molecular Biosciences, Karl-Franzens-University Graz, Humboldtstrasse 50/III, 8010 Graz, Styria, Austria. nikolaus.goessweiner-mohr@uni-graz.at
authors: Nikolaus
authors: N
authors: Goessweiner-Mohr
authors: Lukas
authors: L
authors: Grumet
authors: Tea
authors: T
authors: Pavkov-Keller
authors: Ruth
authors: R
authors: Birner-Gruenberger
authors: Elisabeth
authors: E
authors: Grohmann
authors: Walter
authors: W
authors: Keller
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002021:Buffers; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003227:Conjugation, Genetic; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006094:Gram-Positive Bacteria / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D002021:Buffers; C074281:TraM protein, bacterial
keywords: Gram-positive; TraM; conjugative plasmid transfer; pIP501
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