Surface-layer protein from Caulobacter crescentus: expression, purification and X-ray crystallographic analysis.
Protein surface layers are self-assembling, paracrystalline lattices on the surface of many prokaryotes. Surface-layer proteins have not benefited from widespread structural analysis owing to their resistance to crystallization. Here, the successful expression of a truncated version of RsaA, the surface-layer protein from Caulobacter crescentus, from a Caulobacter protein-expression system is reported. The purification, crystallization and initial X-ray diffraction analysis of the truncated RsaA, the largest surface-layer protein studied to date and the first from a Gram-negative bacterium, are also reported.
Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
Michael D
MD
Jones
Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
Anson C K
AC
Chan
Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
John F
JF
Nomellini
Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
Michael E P
ME
Murphy
Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
John
J
Smit
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016935:Caulobacter crescentus / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014961:X-Ray Diffraction
D001425:Bacterial Outer Membrane Proteins; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C055764:S-layer proteins
Gram-negative; RTX motif; S-layer; phasing; surface-layer protein