The isolation and partial characterization of alpha 2-macroglobulin from the serum of the ostrich (Struthio camelus.
1. alpha 2-Macroglobulin (alpha 2M) activity is present in the serum of the ostrich, Struthio camelus. The chromogenic synthetic peptide substrates BAPNA and ATNA were hydrolysed by trypsin and chymotrypsin, respectively, in the presence of ostrich serum and the alpha 2 M in ostrich serum protected trypsin from being inhibited by soybean trypsin inhibitor. Ostrich alpha 2M proved to be a potent inhibitor of bovine pancreatic trypsin and chymotrypsin. 2. alpha 2M was purified to apparent homogeneity by PEG precipitation, DEAE-Toyopearl 650M, Bio-Gel A-5m and Zn(2+)-affinity chromatography. 3. Ostrich alpha 2M migrated as a single band (M(r) 779,000 during non-denaturing gradient gel electrophoresis and showed increased mobility after reaction with trypsin. Denaturation dissociated ostrich alpha 2M into half-molecules. Denaturation with reduction further dissociated the protein into quarter-subunits. 4. Isoelectric focusing revealed a pI of 5.3. 5. The amino acid composition of ostrich alpha 2M is typical of an alpha 2M, comparing favourably with those of other animal species. The carbohydrate composition of the purified protein, in percentage dry weight of the molecule, was galactose: mannose (1:1), 4.55; N-acetylglucosamine 2.35; N-acetylneuraminic acid, 0.58; and fucose, 0.77. 6. alpha 2M was assessed immunologically by Ouchterlony double-diffusion and Western blot analysis with polyvalent antisera directed against ostrich alpha 2M. 7. Ostrich alpha 2M seems to show many physical, chemical and kinetic properties similar to those of other known alpha 2M(s), but is expected to differ from other alpha Ms when considering the primary structure of the bait region, the area differing among alpha Ms from different species and determining its specificity.
Van Jaarsveld
F
F
Department of Biochemistry, University of Port Elizabeth, South Africa.
Naudé
R J
RJ
Oelofsen
W
W
Travis
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001717:Birds / Q000097:blood*; D007448:Invertebrates / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D013045:Species Specificity; D014714:Vertebrates / Q000097:blood; D000511:alpha-Macroglobulins / Q000737:chemistry / Q000302:isolation & purification*
D000511:alpha-Macroglobulins