CONFORMATIONAL TRANSITION BETWEEN NATIVE AND REACTIVE CENTER CLEAVED FORMS OF ALPHA-1-ANTITRYPSIN BY FOURIER-TRANSFORM INFRARED-SPECTROSCOPY AND SMALL-ANGLE NEUTRON-SCATTERING

α1 Antitrypsin (α1-AT) is the best-characterized member of the serpin superfamily of plasma proteins. Protease inhibitor members of this family undergo a characteristic reactive-center cleavage during expression of their inhibitory activity. The physical basis of this transition in α1-AT from the stressed native conformation to the more stable reactive center cleaved (split) form was studied by Fourier transform infrared (FT-IR) spectroscopy and neutron scattering. The FT-IR spectra show that, while split α1-AT has three intense well-resolved components associated with the presence of antiparallel β-sheet and α-helix conformations, the amide I band of native α1-AT has only one intense component, associated with the presence of β-sheet structure. 1H-2H exchange within the polypeptide backbone, studied by FT-IR and NMR spectroscopy, shows that the native form undergoes greater exchange than the split form. Under the same conditions, neutron scattering shows no differences in the radius of gyration RG of the native and the split forms. In contrast, in high concentrations of phosphate approaching those used for crystallization, the native form (unlike the split form) undergoes dimerization. These data indicate that the conformational transition largely involves localized secondary and tertiary structure rearrangements. We propose that the energetically stressed native α1-AT structure is the consequence of a significantly reduced number of hydrogen bonds in secondary structure components and that reactive-site cleavage between Met358 and Ser359 is the key for the development of the fully hydrogen bonded more stable serpin structure. © 1990, American Chemical Society. All rights reserved.