Altered specificities of genetically engineered {alpha}1 antitrypsin variants |
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Authors: | Jallat S; Carvallo D; Tessier LH; Roecklin D; Roitsch C; Ogushi F; Crystal RG; Courtney M |
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Affiliation: | Department of Molecular Biology, Transgene S A, II rue de Molsheim 67000 Strasbourg, France
1Department of Protein Chemistry, Transgene S A, II rue de Molsheim 67000 Strasbourg, France
2National Heart, Lung and Blood Institute. National Institutes of Health Bethesda, MD 20205, USA |
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Abstract: | Seven active site variants of human 1-antitrypsin ( 1AT) wereproduced in Escherichia coli following site-specific mutagenesisof the 1AT complementary DNA. 1AT (Ala 358), 1AT (Ile358 and 1AT (Val358), were efficient inhibitors of both neutrophil andpancreatic elastases, but not of cathepsin G. 1AT (Ala358, Val358)and 1AT (Phe358 specifically inhibited pancreatic elastase andcathepsin G respectively. The most potent inhibitor of neutrophilelastase was 1AT (Leu358), which also proved to be effectiveagainst cathepsin G. The 1AT (Arg358) variant inactivated thrombinwith kinetics similar to antithrombin III in the presence ofheparin. Electrophoretic analysis showed that SDS-stable highmol. wt complexes were formed between the mutant inhibitorsand the cognate proteases in each case. These data indicatethat effective inhibition occurs when the 1AT P1 residue (position358) corresponds to the primary specificity of the target protease.Moreover, alteration of the P3 residue (position 356) can furthermodify the reactivity of the inhibitor. Two of the variantshave therapeutic potential: 1AT (Leu358 may be more useful thanplasma 1AT in the treatment of destructive lung disorders and 1 (Arg358 could be effective in the control of thrombosis. |
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Keywords: | 1-antitrypsin/" target="_blank">gif" ALT="{alpha}" BORDER="0">1-antitrypsin/ neutrophil proteases/ pulmonary emphysema/ thrombin/ thrombosis |
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