Unfolding studies of human adenovirus type 2 fibre trimers. Evidence for a stable domain.
Adenovirus fibres are trimeric proteins that protrude from the 12 fivefold vertices of the virion and are the cell attachment organelle of the virus. They consist of three segments: an N-terminal tail, which is noncovalently attached to the penton base, a thin shaft carrying 15 amino acid pseudo repeats, and a C-terminal globular head (or knob) which recognizes the primary cell receptor. Due to their exceptional stability, which allows easy distinction of native trimers from unfolded forms and folding intermediates, adenovirus fibres are a very good model system for studying folding in vivo and in vitro. To understand the folding and stability of the trimeric fibres, the unfolding pathway of adenovirus 2 fibres induced by SDS and temperature has been investigated. Unfolding starts from the N-terminus and a stable intermediate accumulates that has the C-terminal head and part of the shaft structure (shown by electron microscopy). The unfolded part can be digested away using limited proteolysis, and the precise digestion sites have been determined. The remaining structured fragment is recognized by monoclonal antibodies that are specific for the trimeric globular head and therefore retains a native trimeric structure. Taken together, our results indicate that adenovirus fibres carry a stable C-terminal domain, consisting of the knob with five shaft-repeats.
Mitraki
A
A
Institut de Biologie Structurale, Grenoble, France. mitraki@ibs.fr
Barge
A
A
Chroboczek
J
J
Andrieu
J P
JP
Gagnon
J
J
Ruigrok
R W
RW
D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry*; D036022:Capsid Proteins; D002918:Chymotrypsin; D006801:Humans; D007700:Kinetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013696:Temperature
D036022:Capsid Proteins; D010446:Peptide Fragments; C018315:hexon capsid protein, Adenovirus; D012967:Sodium Dodecyl Sulfate; D002918:Chymotrypsin