Cloning of cDNA for proteinase 3: a serine protease, antibiotic, and autoantigen from human neutrophils.
Closely similar but nonidentical NH2-terminal amino acid sequences have been reported for a protein or proteins in human neutrophils whose bioactivities is/are diverse (as a serine protease, antibiotic, and Wegener's granulomatosis autoantigen) but that share(s) several features: localization in the azurophil granules, a molecular mass of approximately 29 kD, reactivity with diisopropylfluorophosphate, and the ability to degrade elastin. We previously purified one such entity, termed p29b. Using a monospecific antibody, we have cloned from human bone marrow a cDNA encoding the complete p29b protein in its mature form, along with pre- and pro-sequences. The predicted amino acid sequence agrees closely with the NH2-terminal sequence obtained previously from purified p29b, as well as with sequences newly obtained from CNBr fragments. The primary structure is highly homologous to elastase, cathepsin G, T cell granzymes, and other serine proteases, and shares both the catalytic triad and substrate binding pocket of elastase. Hybridization of the full-length cDNA with restriction enzyme digests of human genomic DNA revealed only one fragment. This suggests that the closely related species described previously are the same, and can be subsumed by the term used for the first-described activity, proteinase 3. Proteinase 3 is more abundant in neutrophils than elastase and has a similar proteolytic profile and specific activity. Thus, proteinase 3 may share the role previously attributed to neutrophil elastase in tissue damage, and has the potential to function as an antimicrobial agent.
Campanelli
D
D
Beatrice and Samuel A. Seaver Laboratory, Department of Medicine, Cornell University Medical College, New York, New York 10021.
Melchior
M
M
Fu
Y
Y
Nakata
M
M
Shuman
H
H
Nathan
C
C
Gabay
J E
JE
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents; D001324:Autoantigens; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000097:blood / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D053146:Myeloblastin; D009504:Neutrophils / Q000201:enzymology* / Q000276:immunology; D010446:Peptide Fragments / Q000302:isolation & purification; D011487:Protein Conformation; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000235:genetics*
D000900:Anti-Bacterial Agents; D001324:Autoantigens; D010446:Peptide Fragments; D004247:DNA; D012697:Serine Endopeptidases; D053146:Myeloblastin