Use of an in-house approach to study the three-dimensional structures of various outer membrane proteins: structure of the alcaligin outer membrane transporter FauA from Bordetella pertussis.
Bordetella pertussis is the bacterial agent of whooping cough in humans. Under iron-limiting conditions, it produces the siderophore alcaligin. Released to the extracellular environment, alcaligin chelates iron, which is then taken up as a ferric alcaligin complex via the FauA outer membrane transporter. FauA belongs to a family of TonB-dependent outer membrane transporters that function using energy derived from the proton motive force. Using an in-house protocol for membrane-protein expression, purification and crystallization, FauA was crystallized in its apo form together with three other TonB-dependent transporters from different organisms. Here, the protocol used to study FauA is described and its three-dimensional structure determined at 2.3 A resolution is discussed.
Brillet
Karl
K
Institut Gilbert-Laustriat, UMR7175 CNRS/Université Louis Pasteur, Strasbourg I, France.
Meksem
Ahmed
A
Lauber
Emmanuelle
E
Reimmann
Cornelia
C
Cobessi
David
D
D001059:Apoproteins / Q000737:chemistry; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D001886:Bordetella pertussis / Q000737:chemistry*; D018360:Crystallography, X-Ray; D006877:Hydroxamic Acids / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D008958:Models, Molecular; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D011522:Protons; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D001059:Apoproteins; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; C121790:FauA protein, Bordetella pertussis; D006877:Hydroxamic Acids; D008565:Membrane Proteins; D011522:Protons; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C092056:alcaligin; C057616:tonB protein, Bacteria; D007501:Iron