Immunological cross-reactivity between human tripeptidyl peptidase II and fibronectin.
Tripeptidyl peptidase II (TPP II) is a large intracellular exopeptidase with an active site of the subtilisin type. Affinity-purified hen antibodies against human erythrocyte TPP II cross-reacted with fibronectin in an immunoblot analysis. Furthermore, antibodies against human fibronectin cross-reacted with TPP II. Antibodies against a 65 kDa cell-binding fragment of fibronectin specifically reacted with TPP II, whereas antibodies against the collagen-binding domain, the main heparin-binding domain or the N-terminal fibrin-binding domain did not react. Moreover, the affinity-purified antibodies against TPP II reacted with a 105 kDa cell-binding fragment of fibronectin but not with the fibrin-binding domain or the collagen-binding domain. When native TPP II was dissociated into smaller units through dialysis against a dilute Tris buffer, it could be digested by chymotrypsin into three stable fragments of 70 kDa, 42 kDa and 20 kDa. It could be demonstrated that the 42 kDa fragment was specifically recognized by antibodies against the 65 kDa cell-binding fragment of fibronectin. Furthermore, labelling with di-[3H]isopropyl phosphorofluoridate and N-terminal sequence determination showed that the 70 kDa fragment contained the active-site serine residue. In conclusion, our findings suggest that one domain of the TPP II molecule bears structural resemblance to a cell-binding fragment of fibronectin.
Tomkinson
B
B
Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
Zetterqvist
O
O
D000595:Amino Acid Sequence; D000626:Aminopeptidases; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D001665:Binding Sites; D002918:Chymotrypsin / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D000939:Epitopes / Q000276:immunology; D005337:Fibrin / Q000378:metabolism; D005353:Fibronectins / Q000276:immunology*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000276:immunology; D012694:Serine; D012697:Serine Endopeptidases / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D005353:Fibronectins; D010446:Peptide Fragments; D012694:Serine; D005337:Fibrin; D006493:Heparin; D003094:Collagen; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C048001:tripeptidyl-peptidase 2; D012697:Serine Endopeptidases; D002918:Chymotrypsin