title: Conformational transition between native and reactive center cleaved forms of alpha 1-antitrypsin by Fourier transform infrared spectroscopy and small-angle neutron scattering.
abstract: alpha 1-Antitrypsin (alpha 1-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 alpha 1-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 alpha 1-AT has three intense well-resolved components associated with the presence of antiparallel beta-sheet and alpha-helix conformations, the amide I band of native alpha 1-AT has only one intense component, associated with the presence of beta-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 alpha 1-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.
authors.affiliation: Department of Protein and Molecular Biology, Royal Free Hospital School of Medicine, London, U.K.
authors.forename: P I
authors.initials: PI
authors.lastname: Haris
authors.forename: D
authors.initials: D
authors.lastname: Chapman
authors.forename: R A
authors.initials: RA
authors.lastname: Harrison
authors.forename: K F
authors.initials: KF
authors.lastname: Smith
authors.forename: S J
authors.initials: SJ
authors.lastname: Perkins
mesh_terms: D005583:Fourier Analysis; D009682:Magnetic Resonance Spectroscopy; D009502:Neutrons; D011487:Protein Conformation; D012542:Scattering, Radiation; D013055:Spectrophotometry, Infrared; D000515:alpha 1-Antitrypsin
chemical_list: D000515:alpha 1-Antitrypsin