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Nearly complete 1H, 13C and 15N chemical shift assignment of monomeric form of N‑terminal domain of Nephila clavipes major ampullate spidroin 2 Nur Alia Oktaviani1 · Ali D. Malay1 · Akimasa Matsugami2 · Fumiaki Hayashi2 · Keiji Numata1,3  Received: 27 June 2020 / Accepted: 31 July 2020 / Published online: 6 August 2020 © Springer Nature B.V. 2020

Abstract Spider dragline silk is well recognized due to its excellent mechanical properties. Dragline silk protein mainly consists of two proteins, namely, major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2). The MaSp N-terminal domain (NTD) conformation displays a strong dependence on ion and pH gradients, which is crucial for the self-assembly behavior of spider silk. In the spider major ampullate gland, where the pH is neutral and concentration of NaCl is high, the NTD forms a monomer. In contrast, within the spinning duct, where pH becomes more acidic (to pH ~ 5) and the concentration of salt is low, NTD forms a dimer in antiparallel orientation. In this study, we report near-complete backbone and side chain chemical shift assignment of the monomeric form of NTD of MaSp2 from Nephila clavipes at pH 7 in the presence of 300 mM NaCl. Our NMR data demonstrate that secondary structure of monomeric form of NTD MaSp2 consists of five helix regions. Keywords  Spider silk · N-terminal domain · NMR · Assignment · Major ampullate spidroin 2 · Nephila clavipes

Biological context Female orb weaving spider can produce up to seven types of silk to perform essential biological tasks, such as prey wrapping, web construction, life line and eggs protection (Gosline et al. 1986; Holland et al. 2019). Among all types of spider silk, spider dragline silk has been the focus of many studies due to its large diameter size, accessibility * Nur Alia Oktaviani [email protected] * Keiji Numata [email protected] 1



Biomacromolecules Research Team, RIKEN Center for Sustainable Resource Science, 2‑1 Hirosawa, Wako, Saitama 351‑0198, Japan

2



Advanced NMR Application and Platform Team, NMR Research and Collaboration Group, NMR Science and Development Division, RIKEN SPring-8 Center, 1‑7‑22 Suehiro‑cho, Tsurumi‑ku, Yokohama, Kanagawa 230‑0045, Japan

3

Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Kyoto‑Daigaku‑Katsura, Nishikyo‑ku, Kyoto 615‑8510, Japan



and mechanical properties (Xu and Lewis 1990). Based on an equal basis weight, spider dragline silk exhibits higher toughness than steel and Kevlar (Vepari and Kaplan 2007; Heim et al. 2009), which make it promising for industrial and biomedical applications (Altman et al. 2003). Spider dragline silk of Nephila clavipes is primarily composed of major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2) (Xu and Lewis 1990; Hinman and Lewis 1992; Sponner et al. 2005; Malay et al. 2017). Both MaSp1 and MaSp2 consist of conserved N-terminal (NTD) and C-terminal (CTD) domains, flanking a long repetitive central domain. In the gland, the spider silk

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