DAF- and collagen-binding properties of chimeric Dr fimbriae.
Display of heterologous proteins on the surface of micro-organisms has become an increasingly used strategy for a range of applications in microbiology, biotechnology and vaccinology. In this study, the potential of the major structural protein DraE of Escherichia coli Dr fimbriae as a display system for heterologous sequences was tested. One copy of a heterologous sequence mimicking a small P(k) epitope of simian virus 5 was inserted into the draE gene, replacing the N-terminal region of the surface-exposed domain 2 as previously done with the glycoprotein D of herpes simplex virus type 1. The exposure of chimeric proteins on the bacterial surface was detected by immunofluorescence microscopy. Insertion of the heterogenic peptides had no detectable effect on the Ig-barrel structure of the DraE fimbrial subunits, as confirmed by FTIR spectroscopy. Additionally, the affinity of the chimeric fimbriae for DAF and type IV collagen was similar to that of the wild-type Dr fimbriae.
Department of Microbiology, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-952 Gdańsk, Poland.
Beata
B
Zalewska
Department of Physical Chemistry, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-952 Gdańsk, Poland.
Janusz
J
Stangret
Department of Microbiology, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-952 Gdańsk, Poland.
Katarzyna
K
Bury
Department of Pharmaceutical Technology and Biochemistry, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-952 Gdańsk, Poland.
Marek
M
Wojciechowski
Department of Microbiology, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-952 Gdańsk, Poland.
Józef
J
Kur
Department of Microbiology, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-952 Gdańsk, Poland.
Rafał
R
Piątek
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018830:Adhesins, Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003094:Collagen / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D045402:Parainfluenza Virus 5 / Q000235:genetics; D017434:Protein Structure, Tertiary; D011971:Receptors, Immunologic / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared
D018829:Adhesins, Bacterial; D018830:Adhesins, Escherichia coli; C465564:Dr adhesin, E coli; C465563:DraE protein, E coli; D029968:Escherichia coli Proteins; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; C047105:bacterial adhesin receptor; D003094:Collagen