Some properties of the alkaline proteinase from Aspergillus melleus.
Seaprose is a semi-alkaline proteinase produced by Aspergillus melleus. The aim of our study was to further characterize the properties of this enzyme, particularly looking at its interaction with alpha 1-proteinase inhibitor, the major human plasma proteinase inhibitor. We studied the cleavage of three synthetic peptide substrates induced by seaprose and the inhibitory profile of the enzyme by means of a panel of inhibitors, including alpha 1-proteinase inhibitor. The interaction between seaprose and alpha 1-proteinase inhibitor was also studied with SDS-PAGE. Finally, the elastolytic activity of seaprose was checked by means of bovine elastin solubilization. We found that seaprose cleaves preferentially the substrate containing a Phe residue in the P1 position. The inhibitory profile showed that seaprose is a serine-proteinase that cannot be inhibited by alpha 1-proteinase inhibitor. The SDS-PAGE revealed that alpha 1-proteinase inhibitor, after incubation with seaprose, underwent a limited proteolysis. Finally, seaprose 10(-2) M and 10(-3) M was able to solubilize bovine elastin. We conclude that seaprose is a serine-proteinase able to inactivate human alpha 1-proteinase inhibitor with limited proteolysis at (or near) the active site and that it has mild elastinolytic capacity.
Luisetti
M
M
Istituto di Tisiologia e Malattie Respiratoire, IRCCS Policlinico San Matteo, Università di Pavia, Italy.
Piccioni
P D
PD
Dyne
K
K
Donnini
M
M
Bulgheroni
A
A
Pasturenzi
L
L
Donnetta
A M
AM
Peona
V
V
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology*; D004549:Elastin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000378:metabolism*; D012697:Serine Endopeptidases; D012995:Solubility; D000515:alpha 1-Antitrypsin / Q000378:metabolism*
D000515:alpha 1-Antitrypsin; D004549:Elastin; D010447:Peptide Hydrolases; C022624:Seaprose S; D012697:Serine Endopeptidases