Commercial plasma alpha1-antitrypsin (Prolastin) contains a conformationally inactive, latent component.
Fractionated plasma alpha1-antitrypsin is widely-used as replacement therapy in patients with Z alpha1-antitrypsin deficiency-related emphysema. We have recently shown that purified antitrypsin may be induced to adopt an inactive latent conformation by heating at high temperatures in stabilizing concentrations of sodium citrate. Such a conformation was predicted to be present in commercial preparations of antitrypsin, as these require heating under similar conditions for viral inactivation. Native antitrypsin was purified from plasma, and commercial antitrypsin (Prolastin) was obtained from Bayer Corporation. Western blot analysis of transverse urea gradient (TUG) gels showed that commercial antitrypsin migrated as two bands: one with an unfolding profile of native antitrypsin and the second with a profile of latent antitrypsin. A latent fraction, comprising approximately 8% of the total antitrypsin, was separated from the native antitrypsin in Prolastin by anion exchange chromatography. The specific activity of this latent form against bovine alpha-chymotrypsin increased from 1 to 2% to 50% over 3 h after refolding from 6 M guanidine hydrochloride. These data show that commercial antitrypsin contains a latent component. The significance of this conformation in vivo is unknown, although Prolastin has shown few adverse side-effects in prolonged clinical usage.
Lomas
D A
DA
Dept of Medicine, University of Cambridge, MRC Centre, UK.
Elliott
P R
PR
Carrell
R W
RW
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010641:Phenotype; D011487:Protein Conformation; D011656:Pulmonary Emphysema / Q000188:drug therapy; D015842:Serine Proteinase Inhibitors / Q000737:chemistry* / Q000172:deficiency / Q000627:therapeutic use; D000515:alpha 1-Antitrypsin / Q000737:chemistry* / Q000627:therapeutic use; D019896:alpha 1-Antitrypsin Deficiency
D015842:Serine Proteinase Inhibitors; D000515:alpha 1-Antitrypsin; D002918:Chymotrypsin