Isolation and characterization of alpha-1-proteinase inhibitor from goat plasma.
Alpha-1-proteinase inhibitor (alpha-1-PI) was isolated from goat plasma by salt fractionation, and chromatography on a DEAE-cellulose column. The inhibitor was found to be homogeneous by gel chromatography, SDS-PAGE and PAGE.Mr values by gel filtration (57 kDa), and by SDS-PAGE (52 kDa), under reducing conditions were nearly the same suggesting that the inhibitor consists of a single polypeptide chain. It contained 13.8% neutral hexose but no sialic acid residue. The values of isoionic pH, and extinction coefficient at 278 nm were 4.84, and 4.6, respectively. Fluorescence spectral properties showed tryptophan residues in the inhibitor. Solvent perturbation difference spectra suggested 74% exposure of the tryptophan residues in the native molecule. Gel filtration behaviour of the inhibitor was consistent with a Stokes radius of 3.16 nm, diffusion coefficient of 7.02 X 10(-7) cm2-sec-1 and a frictional ratio of 1.24 suggesting asymmetry and/or excessive hydration of the inhibitor molecule. Goat alpha-1-PI, unlike human alpha-1-PI was found to be potent inhibitor of bovine trypsin but a poor inhibitor of porcine pancreatic elastase. It was virtually devoid of antichymotryptic activity.
Department of Biochemistry, J.N.M.C., Aligarh, India.
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Salahuddin
D000818:Animals; D005591:Chemical Fractionation; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D004591:Electrophoresis, Polyacrylamide Gel; D006041:Goats / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors; D010455:Peptides / Q000032:analysis; D013050:Spectrometry, Fluorescence; D014357:Trypsin; D014361:Trypsin Inhibitors; D014364:Tryptophan / Q000032:analysis; D000515:alpha 1-Antitrypsin / Q000737:chemistry* / Q000302:isolation & purification
D010455:Peptides; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin; D014364:Tryptophan; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D014357:Trypsin