The isolation and partial characterization of alpha 1-proteinase inhibitor from the serum of the ostrich (Struthio camelus).
1. Native and cleaved alpha 1-proteinase inhibitor was purified from ostrich serum using Sepharose-blue dextran chromatography, ammonium sulfate precipitation and ion exchange chromatography on DEAE-Toyopearl 650 M at pH 8.8 and 6.5. Ostrich alpha 1-PI displayed M(r) values of 68,100 using gradient PAGE and 66,200 using Ferguson plots. Isoelectric focusing of ostrich alpha 1-PI in the pH range 3-10 revealed pI values of 4.84 and 4.91, and in the pH range 4-6 the characteristic microheterogeneity observed for mammalian alpha 1-PIs was displayed. The presence of sialic acid, hexoses and hexosamines was detected using chemical methods, but were found in much lower quantities as compared to alpha 1-PIs of other species. Western blot analysis demonstrated a positive reaction between the native and cleaved ostrich alpha 1-PIs and the antibodies to the ostrich alpha 1-PIs raised in rabbits. No cross-reactivity was demonstrated by Western blot analysis between human alpha 1-PI and antibodies to ostrich alpha 1-PI. The inhibitory effect of alpha 1-PI on elastase and chymotrypsin was also investigated.
Kuhn
C R
CR
Department of Biochemistry, University of Port Elizabeth, South Africa.
Naudé
R J
RJ
Travis
J
J
Oelofsen
W
W
D000595:Amino Acid Sequence; D000818:Animals; D001717:Birds / Q000097:blood*; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D006595:Hexosamines / Q000032:analysis; D006601:Hexoses / Q000032:analysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors; D017386:Sequence Homology, Amino Acid; D012794:Sialic Acids / Q000032:analysis; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology
D006595:Hexosamines; D006601:Hexoses; D012794:Sialic Acids; D000515:alpha 1-Antitrypsin; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D019158:N-Acetylneuraminic Acid