title: The structural basis of serpin polymerization studied by hydrogen/deuterium exchange and mass spectrometry.
abstract: The serpinopathies are a group of inherited disorders that share as their molecular basis the misfolding and polymerization of serpins, an important class of protease inhibitors. Depending on the identity of the serpin, conditions arising from polymerization include emphysema, thrombosis, and dementia. The structure of serpin polymers is thus of considerable medical interest. Wild-type alpha(1)-antitrypsin will form polymers upon incubation at moderate temperatures and has been widely used as a model system for studying serpin polymerization. Using hydrogen/deuterium exchange and mass spectrometry, we have obtained molecular level structural information on the alpha(1)-antitrypsin polymer. We found that the flexible reactive center loop becomes strongly protected upon polymerization. We also found significant increases in protection in the center of beta-sheet A and in helix F. These results support a model in which linkage between serpins is achieved through insertion of the reactive center loop of one serpin into beta-sheet A of another. We have also examined the heat-induced conformational changes preceding polymerization. We found that polymerization is preceded by significant destabilization of beta-sheet C. On the basis of our results, we propose a mechanism for polymerization in which beta-strand 1C is displaced from the rest of beta-sheet C through a binary serpin/serpin interaction. Displacement of strand 1C triggers further conformational changes, including the opening of beta-sheet A, and allows for subsequent polymerization.
authors.affiliation: Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, OH 44106, USA.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Tsutsui
authors.forename: Barbara
authors.initials: B
authors.lastname: Kuri
authors.forename: Tanusree
authors.initials: T
authors.lastname: Sengupta
authors.forename: Patrick L
authors.initials: PL
authors.lastname: Wintrode
mesh_terms: D020134:Catalytic Domain / Q000502:physiology; D041961:Deuterium Exchange Measurement / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry; D008958:Models, Molecular; D020836:Protein Structure, Quaternary / Q000502:physiology; D017433:Protein Structure, Secondary / Q000502:physiology; D000515:alpha 1-Antitrypsin / Q000737:chemistry* / Q000378:metabolism; D019896:alpha 1-Antitrypsin Deficiency / Q000378:metabolism
chemical_list: D000515:alpha 1-Antitrypsin
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