title: Novel assembly properties of recombinant spider dragline silk proteins.
abstract: Spider dragline silk, which exhibits extraordinary strength and toughness, is primarily composed of two related proteins that largely consist of repetitive sequences. In most spiders, the repetitive region of one of these proteins is rich in prolines, which are not present in the repetitive region of the other. The absence of prolines in one component was previously speculated to be essential for the thread structure. Here, we analyzed dragline proteins of the garden spider Araneus diadematus, ADF-3 and ADF-4, which are both proline rich, by employing the baculovirus expression system. Whereas ADF-3 represented an intrinsically soluble protein, ADF-4 was insoluble in vitro and self-assembled into filaments in the cytosol of the host insect cells. These ADF-4 filaments displayed the exceptional chemical stability of authentic silk threads. We provide evidence that the observed properties of ADF-3 and ADF-4 strongly depend on intrinsic characteristics such as hydropathicity, which differs dramatically between the two proteins, as in most other pairs of dragline silk proteins from other Araneoidea species, but not on their proline content. Our findings shed new light on the structural components of spider dragline silk, allowing further elucidation of their assembly properties, which may open the door for commercial applications.
authors.affiliation: Department of Organic Chemistry and Biochemistry, Technische Universität München, 85747 Garching, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Huemmerich
authors.forename: Thomas
authors.initials: T
authors.lastname: Scheibel
authors.forename: Fritz
authors.initials: F
authors.lastname: Vollrath
authors.forename: Shulamit
authors.initials: S
authors.lastname: Cohen
authors.forename: Uri
authors.initials: U
authors.lastname: Gat
authors.forename: Shmulik
authors.initials: S
authors.lastname: Ittah
mesh_terms: D000818:Animals; D016367:Baculoviridae; D005349:Fibroins / Q000096:biosynthesis* / Q000737:chemistry / Q000648:ultrastructure; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D010957:Plasmids / Q000235:genetics; D011392:Proline / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D013112:Spiders / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D005349:Fibroins; D011392:Proline