Proteolytic sensitivity and helper T-cell epitope immunodominance associated with the mobile loop in Hsp10s.
Antigen three-dimensional structure potentially limits antigen processing and presentation to helper T-cell epitopes. The association of helper T-cell epitopes with the mobile loop in Hsp10s from mycobacteria and bacteriophage T4 suggests that the mobile loop facilitates proteolytic processing and presentation of adjacent sequences. Sites of initial proteolytic cleavage were mapped in divergent Hsp10s after treatment with a variety of proteases including cathepsin S. Each protease preferentially cleaved the Hsp10s in the mobile loop. Flexibility in the 22-residue mobile loop most probably allows it to conform to protease active sites. Three variants of the bacteriophage T4 Hsp10 were constructed with deletions in the mobile loop to test the hypothesis that shorter loops would be less sensitive to proteolysis. The two largest deletions effectively inhibited proteolysis by several proteases. Circular dichroism spectra and chemical cross-linking of the deletion variants indicate that the secondary and quaternary structures of the variants are native-like, and all three variants were more thermostable than the wild-type Hsp10. Local structural flexibility appears to be a general requirement for proteolytic sensitivity, and thus, it could be an important factor in antigen processing and helper T-cell epitope immunogenicity.
Carmicle
Stephanie
S
Department of Biochemistry, Tulane University Health Sciences Center, New Orleans, Louisiana 70112-2699, USA.
Dai
Guixiang
G
Steede
N Kalaya
NK
Landry
Samuel J
SJ
D000595:Amino Acid Sequence; D017951:Antigen Presentation; D018835:Chaperonin 10 / Q000737:chemistry* / Q000276:immunology; D018984:Epitopes, T-Lymphocyte; D006801:Humans; D016056:Immunodominant Epitopes; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*
D018835:Chaperonin 10; D018984:Epitopes, T-Lymphocyte; D016056:Immunodominant Epitopes