title: Native-sized spider silk proteins synthesized in planta via intein-based multimerization.
abstract: The synthesis of native-sized proteins is a pre-requisite for exploiting the potential of spider silk as a bio-based material. The unique properties of spider silk, such as extraordinary tensile strength and elasticity, result from the highly repetitive nature of spider silk protein motifs. The present report describes the combination of spider silk flagelliform protein (FLAG) production in the endoplasmic reticulum of tobacco plant leaf cells with an intein-based posttranslational protein fusion technology. The repeated ligation of FLAG monomers resulted in the formation of large multimers. This method avoids the need for highly repetitive transgenes, which may result in a higher genetic and transcriptional stability. Here we show, for the first time, the production of synthetic, high molecular weight spider silk proteins larger than 250 kDa based on the assembly of protein monomers via intein-mediated trans-splicing in planta. The resulting multimeric structures form microfibers, thereby demonstrating their great potential as a biomaterial.
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, 06466 Stadt Seeland, OT Gatersleben, Germany.
authors.forename: Valeska
authors.initials: V
authors.lastname: Hauptmann
authors.forename: Nicola
authors.initials: N
authors.lastname: Weichert
authors.forename: Matthias
authors.initials: M
authors.lastname: Menzel
authors.forename: Dominic
authors.initials: D
authors.lastname: Knoch
authors.forename: Norman
authors.initials: N
authors.lastname: Paege
authors.forename: Jürgen
authors.initials: J
authors.lastname: Scheller
authors.forename: Uwe
authors.initials: U
authors.lastname: Spohn
authors.forename: Udo
authors.initials: U
authors.lastname: Conrad
authors.forename: Mario
authors.initials: M
authors.lastname: Gils
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000096:biosynthesis / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D019476:Insect Proteins; D047668:Inteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D055503:Protein Multimerization; D047011:Silk / Q000235:genetics; D013112:Spiders / Q000737:chemistry / Q000235:genetics; D014026:Nicotiana / Q000235:genetics*; D020040:Trans-Splicing / Q000235:genetics
chemical_list: D060829:Arthropod Proteins; D019476:Insect Proteins; D047011:Silk