title: The structure of a GFP-based antibody (fluorobody) to TLH, a toxin from Vibrio parahaemolyticus.
abstract: A fluorobody is a manmade hybrid molecule that is composed of green fluorescent protein (GFP) and a fragment of antibody, which combines the affinity and specificity of an antibody with the visibility of a GFP. It is able to provide a real-time indication of binding while avoiding the use of tags and secondary binding reagents. Here, the expression, purification and crystal structure of a recombinant fluorobody for TLH (thermolabile haemolysin), a toxin from the lethal food-borne disease bacterium Vibrio parahaemolyticus, are presented. This is the first structure of a fluorobody to be reported. Crystals belonging to space group P4(3)2(1)2, with unit-cell parameters a = b = 63.35, c = 125.90 Å, were obtained by vapour diffusion in hanging drops and the structure was refined to an Rfree of 16.7% at 1.5 Å resolution. The structure shows a CDR loop of the antibody on the GFP scaffold.
authors.affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, 155 Yangqiao Road West, Fuzhou 350002, People's Republic of China.
authors.forename: Yaoguang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, 155 Yangqiao Road West, Fuzhou 350002, People's Republic of China.
authors.forename: Xiaocheng
authors.initials: X
authors.lastname: Huang
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, People's Republic of China.
authors.forename: Rongzhi
authors.initials: R
authors.lastname: Wang
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, People's Republic of China.
authors.forename: Shihua
authors.initials: S
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, 155 Yangqiao Road West, Fuzhou 350002, People's Republic of China.
authors.forename: Ning
authors.initials: N
authors.lastname: Shi
mesh_terms: D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D014736:Vibrio parahaemolyticus
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D006460:Hemolysin Proteins; C073845:hemolysin, Vibrio; D049452:Green Fluorescent Proteins
keywords: CDR3; GFP scaffold; TLH; fluorobody