Formation of highly stable chimeric trimers by fusion of an adenovirus fiber shaft fragment with the foldon domain of bacteriophage t4 fibritin.
The folding of beta-structured, fibrous proteins is a largely unexplored area. A class of such proteins is used by viruses as adhesins, and recent studies revealed novel beta-structured motifs for them. We have been studying the folding and assembly of adenovirus fibers that consist of a globular C-terminal domain, a central fibrous shaft, and an N-terminal part that attaches to the viral capsid. The globular C-terminal, or "head" domain, has been postulated to be necessary for the trimerization of the fiber and might act as a registration signal that directs its correct folding and assembly. In this work, we replaced the head of the fiber by the trimerization domain of the bacteriophage T4 fibritin, termed "foldon." Two chimeric proteins, comprising the foldon domain connected at the C-terminal end of four fiber shaft repeats with or without the use of a natural linker sequence, fold into highly stable, SDS-resistant trimers. The structural signatures of the chimeric proteins as seen by CD and infrared spectroscopy are reported. The results suggest that the foldon domain can successfully replace the fiber head domain in ensuring correct trimerization of the shaft sequences. Biological implications and implications for engineering highly stable, beta-structured nanorods are discussed.
Papanikolopoulou
Katerina
K
Institut de Biologie Structurale, 41 rue Jules Horowitz, Grenoble 38027, France.
Forge
Vincent
V
Goeltz
Pierrette
P
Mitraki
Anna
A
D036022:Capsid Proteins / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D015202:Protein Engineering; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry
D036022:Capsid Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C057839:fibritin protein, Enterobacteria phage T4; C018315:hexon capsid protein, Adenovirus