No abstract

Spiders synthesize several kinds of silk fibers. In the primary structure of spider silk, one of the major ampullate (dragline, frame) silks, spidroin 1, and flagelliform silk (core fibers of adhesive spiral), there are common repeated X-Gly-Gly (X = Ala, Leu, Pro, Tyr, Glu, and Arg) sequences, which are considered to be related to the elastic character of these fibers. In this paper, two dimensional spin diffusion solid-state NMR under off magic angle spinning (OMAS), (13)C chemical shift contour plots, and Rotational Echo DOuble Resonance (REDOR) were applied to determine the torsion angles of one Ala and two kinds of Gly residues in the Ala-Gly-Gly sequence of (13)C=O isotope-labeled (Ala-Gly-Gly)(10). The torsion angles were determined to be (phi, psi) = (-90 degrees, 150 degrees ) within an experimental error of +/-10 degrees for each residue. This conformation is characterized as 3(1) helix which is in agreement with the structure proposed from the X-ray powder diffraction pattern of poly(Ala-Gly-Gly). The 3(1) helix of (Ala-Gly-Gly)(10) does not change by formic acid treatment although (Ala-Gly)(15) easily changes from the silk I conformation (the structure of Bombyx mori silk fibroin before spinning in the solid state) to silk II conformation (the structure of the silk fiber after spinning) by such treatment. Thus, the 3(1) helix conformation of (Ala-Gly-Gly)(10) is considered very stable. Furthermore, the torsion angles of the 16th Leu residue of (Leu-Gly-Gly)(10) were also determined as (phi, psi) = (-90 degrees, 150 degrees ) and this peptide is also considered to take 3(1) helix conformation.

The structure of an elastin-mimetic model peptide, (VPGVG) 6, was proposed by combining data obtained from quantitative use of the conformation-dependent 13 C NMR chemical shifts, twodimensional off magic angle spinning spin-diffusion solid-state NMR with 13 C double labeling of the peptides, rotational-echo double-resonance of 13 C, 15 N double labeling peptides, and statistical distribution of the backbone torsion angles of Val-Pro, Pro-Gly, and Gly-Val-Gly sequences from PDB. In essence, this approach drew upon several sets of data to formulate the resulting model, namely, that there is a distribution of conformations in this polypeptide. The Val-16 residue adopts torsion angles (φ, ψ) ) (-90 ( 15°, 120 ( 15°). In contrast, bimodal distributions for the central residues of the (VPGVG) subunit were detected. For Pro-12, (φ, ψ) ) (-60 ( 15°, 120 ( 15°), 80%, and (φ, ψ) ) (-60 ( 15°, -30 ( 15°), 20%; for Gly-13, (φ, ψ) ) (90 ( 30°, -15 ( 30°) and (φ, ψ) ) (-90 ( 15°, 0 ( 15°), 60%, and (φ, ψ) ) (-105 ( 75°, 150 ( 30°), (φ, ψ) ) (120 ( 60°, 150 ( 30°), (φ, ψ) ) (-120 ( 60°, -150 ( 30°) and (φ, ψ) ) (120 ( 60°, -150 ( 30°), 40%. For Val-14, (φ, ψ) ) (-110 ( 20°, 130 ( 20°), 70%, and (φ, ψ) ) (-75 ( 15°, -15 ( 15°) 30%. To reconcile the torsion angles corresponding to the main component in the conformational distributions, a type-II β-turn structure was assigned to about 40% of the Pro-Gly pair.