Purification and partial characterisation of alpha(2)-antiplasmin and plasmin(ogen) from ostrich plasma.
This study reports the isolation and partial characterisation of the ostrich serpin, alpha(2)AP, and its target enzyme, ostrich plasmin, in its active and inactive proenzyme, namely plasminogen, forms. Ostrich alpha(2)AP was purified using L-lysine-Sepharose chromatography, ammonium sulfate fractionation, and Super Q-650S and ostrich LBSI-Sepharose chromatographies. It revealed a M(r) of 84 K (thousand) and had one and two N-terminal amino acids in common with 11 of those of human and bovine alpha(2)AP, respectively. It showed the largest inhibitory effect on ostrich plasmin, followed by bovine trypsin and plasmin, respectively, and much less plasmin inhibition than bovine aprotinin, but much more so than human alpha(2)AP, DFP and EACA. Ostrich plasminogen was highly purified after L-lysine-Sepharose chromatography and showed a M(r) of 92 K, a total of 775 amino acids and its N-terminal sequence showed approximately 53% identity with those of human, rabbit, cat, and ox plasminogens. Ostrich plasmin, obtained by the urokinase-activation of ostrich plasminogen, revealed a M(r) of 78 K, a total of 638 amino acids, an N-terminal sequence showing two to four residues identical to five of those of human, cat, dog, rabbit, and ox plasmins, and pH and temperature optima of 8.0 and 40 degrees C, respectively.
Thomas
A R
AR
Department of Biochemistry and Microbiology, University of Port Elizabeth, P.O. Box 1600, 6000, Port Elizabeth, South Africa.
Naudé
R J
RJ
Oelofsen
W
W
Naganuma
T
T
Muramoto
K
K
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D007611:Aprotinin / Q000737:chemistry; D001665:Binding Sites; D002415:Cats; D002417:Cattle; D002847:Chromatography, Agarose; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D010958:Plasminogen / Q000096:biosynthesis* / Q000737:chemistry*; D011817:Rabbits; D015843:Serpins / Q000737:chemistry; D020310:Struthioniformes; D013696:Temperature; D014357:Trypsin / Q000737:chemistry; D000979:alpha-2-Antiplasmin / Q000096:biosynthesis* / Q000737:chemistry*
D000596:Amino Acids; D004792:Enzyme Precursors; D015843:Serpins; D000979:alpha-2-Antiplasmin; D010958:Plasminogen; D007611:Aprotinin; D014357:Trypsin