Purification, crystallization and preliminary X-ray diffraction analysis of an oomycete-derived Nep1-like protein.
The elicitor protein Nep1-like protein from the plant pathogen Pythium aphanidermatum was purified and crystallized using the hanging-drop vapour-diffusion method. A native data set was collected to 1.35 A resolution at 100 K using synchrotron radiation. Since selenomethionine-labelled protein did not crystallize under the original conditions, a second crystal form was identified that yielded crystals that diffracted to 2.1 A resolution. A multiple-wavelength anomalous dispersion (MAD) experiment was performed at 100 K and all four selenium sites were identified, which allowed solution of the structure.
Luberacki
Borries
B
Center for Plant Molecular Biology, Auf der Morgenstelle 5, 72076 Tübingen, Universität Tübingen, Germany.
Weyand
Michael
M
Seitz
Ulrich
U
Koch
Wolfgang
W
Oecking
Claudia
C
Ottmann
Christian
C
D020418:Algal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D011775:Pythium / Q000378:metabolism*; D012645:Selenomethionine / Q000737:chemistry / Q000378:metabolism; D014961:X-Ray Diffraction
D020418:Algal Proteins; D012645:Selenomethionine