title: Structural characterization of Spinacia oleracea trypsin inhibitor III (SOTI-III).
abstract: In recent decades, several canonical serine protease inhibitor families have been classified and characterized. In contrast to most trypsin inhibitors, those from garden four o'clock (Mirabilis jalapa) and spinach (Spinacia oleracea) do not share sequence similarity and have been proposed to form the new Mirabilis serine protease inhibitor family. These 30-40-amino-acid inhibitors possess a defined disulfide-bridge topology and belong to the cystine-knot miniproteins (knottins). To date, no atomic structure of this inhibitor family has been solved. Here, the first structure of S. oleracea trypsin inhibitor III (SOTI-III), in complex with bovine pancreatic trypsin, is reported. The inhibitor was synthesized by solid-phase peptide synthesis on a multi-milligram scale and was assayed to test its inhibitory activity and binding properties. The structure confirmed the proposed cystine-bridge topology. The structural features of SOTI-III suggest that it belongs to a new canonical serine protease inhibitor family with promising properties for use in protein-engineering and medical applications.
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Glotzbach
authors.forename: Stefan
authors.initials: S
authors.lastname: Schmelz
authors.forename: Michael
authors.initials: M
authors.lastname: Reinwarth
authors.forename: Andreas
authors.initials: A
authors.lastname: Christmann
authors.forename: Dirk W
authors.initials: DW
authors.lastname: Heinz
authors.forename: Harald
authors.initials: H
authors.lastname: Kolmar
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000737:chemistry*; D002417:Cattle; D003460:Crystallization; D018360:Crystallography, X-Ray; D039282:Mirabilis / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry* / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D018724:Spinacia oleracea / Q000737:chemistry*
chemical_list: D010940:Plant Proteins; D015842:Serine Proteinase Inhibitors; D007611:Aprotinin