Nonnative interactions between cysteines direct productive assembly of P22 tailspike protein.
Nonnative disulfide bond formation can play a critical role in the assembly of disulfide bonded proteins. During the folding and assembly of the P22 tailspike protein, nonnative disulfide bonds form both in vivo and in vitro. However, the mechanism and identity of cysteine disulfide pairs remains elusive, particularly for P22 tailspike, which contains no disulfide bonds in its native, functional form. Understanding the interactions between cysteine residues is important for developing a mechanistic model for the role of nonnative cysteines in P22 tailspike assembly. Prior in vivo studies have suggested that cysteines 496, 613, and 635 are the most likely site for sulfhydryl reactivity. Here we demonstrate that these three cysteines are critical for efficient assembly of tailspike trimers, and that interactions between cysteine pairs lead to productive assembly of native tailspike.
Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716, USA.
Brenda L
BL
Danek
Anne Skaja
AS
Robinson
D019943:Amino Acid Substitution; D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry*; D019281:Dimerization; D006026:Glycoside Hydrolases; D046911:Macromolecular Substances; D009154:Mutation; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry; D012694:Serine / Q000737:chemistry*; D013329:Structure-Activity Relationship; D017376:Viral Tail Proteins / Q000737:chemistry*
D046911:Macromolecular Substances; D011994:Recombinant Proteins; D017376:Viral Tail Proteins; D012694:Serine; D006026:Glycoside Hydrolases; C083268:tailspike protein, bacteriophage; D003545:Cysteine