Natural processing sites for human cathepsin E and cathepsin D in tetanus toxin: implications for T cell epitope generation.
Cathepsin E is an aspartic proteinase that has been implicated frequently in Ag processing for presentation on class II MHC molecules, but no information exists on its cleavage specificity within Ags in relation to known T cell epitopes. We have analyzed the processing by cathepsin E of a large C-terminal domain of tetanus toxin (residues 872-1315), and we have compared the processing products with those liberated by cathepsin D, a related aspartic proteinase also thought to be involved in class II MHC-restricted Ag processing. Processing products were analyzed by N-terminal Edman degradation and mass spectrometry following reverse-phase HPLC separation of peptides. A total of 28 cleavage sites was identified, 11 of which were recognized by both cathepsins E and D. Most, although not all, sites were between pairs of hydrophobic residues and were located within the 200-amino-acid C terminal region known to contain several human T cell epitopes. Previously described T cell epitopes, for example, between residues 1273 and 1284, were flanked by cathepsin E and D cleavage sites. These data are consistent with an important role for cathepsins E and/or D in Ag processing in the human immune system.
Hewitt
E W
EW
Department of Biochemistry, Medical Sciences Institute, University of Dundee, United Kingdom.
Treumann
A
A
Morrice
N
N
Tatnell
P J
PJ
Kay
J
J
Watts
C
C
D000595:Amino Acid Sequence; D017951:Antigen Presentation; D002402:Cathepsin D / Q000276:immunology* / Q000378:metabolism; D020059:Cathepsin E; D002403:Cathepsins / Q000276:immunology* / Q000378:metabolism; D002999:Clone Cells / Q000276:immunology / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000096:biosynthesis* / Q000276:immunology / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism / Q000502:physiology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000201:enzymology / Q000276:immunology; D013744:Tetanus Toxin / Q000276:immunology* / Q000378:metabolism
D018984:Epitopes, T-Lymphocyte; D010446:Peptide Fragments; D013744:Tetanus Toxin; D002403:Cathepsins; D020059:Cathepsin E; D002402:Cathepsin D