Spider wrapping silk fibre architecture arising from its modular soluble protein precursor

Marie Laurence Tremblay; Lingling Xu; Thierry Lefèvre; Muzaddid Sarker; Kathleen E. Orrell; Jérémie Leclerc; Qing Meng; Michel Pézolet; Michèle Auger; Xiang Qin Liu; Jan K. Rainey

doi:10.1038/srep11502

Spider wrapping silk fibre architecture arising from its modular soluble protein precursor

Marie Laurence Tremblay, Lingling Xu, Thierry Lefèvre, Muzaddid Sarker, Kathleen E. Orrell, Jérémie Leclerc, Qing Meng, Michel Pézolet, Michèle Auger, Xiang Qin Liu, Jan K. Rainey

Medicine

Research output: Contribution to journal › Article › peer-review

45 Citations (Scopus)

Abstract

Spiders store spidroins in their silk glands as high concentration aqueous solutions, spinning these dopes into fibres with outstanding mechanical properties. Aciniform (or wrapping) silk is the toughest spider silk and is devoid of the short amino acid sequence motifs characteristic of the other spidroins. Using solution-state NMR spectroscopy, we demonstrate that the 200 amino acid Argiope trifasciata AcSp1 repeat unit contrasts with previously characterized spidroins, adopting a globular 5-helix bundle flanked by intrinsically disordered N- and C-terminal tails. Split-intein-mediated segmental NMR-active isotope-enrichment allowed unambiguous demonstration of modular and malleable "beads-on-a-string" concatemeric behaviour. Concatemers form fibres upon manual drawing with silk-like morphology and mechanical properties, alongside secondary structuring and orientation consistent with native AcSp1 fibres. AcSp1 structural stability varies locally, with the fifth helix denaturing most readily. The structural transition of aciniform spidroin from a mostly α-helical dope to a mixed α-helix/β-sheet-containing fibre can be directly related to spidroin architecture and stability.

Original language	English
Article number	11502
Journal	Scientific Reports
Volume	5
DOIs	https://doi.org/10.1038/srep11502
Publication status	Published - Jun 26 2015

Bibliographical note

Funding Information:
Thanks to Drs. Stephen Bearne and David Waisman for CD spectropolarimeter access; Bruce Stewart for technical assistance; Xudong Dai for fibre mechanical properties testing; Dr. Mike Lumsden for 11.7 T NMR spectrometer support at the Nuclear Magnetic Resonance Research Resource Facility (NMR3, Dalhosie University); Ian Burton and Drs. Nadine Merkley and Ray Syvitski for 16.4 T NMR spectrometer support at the National Research Council Biological Magnetic Resonance Facility (NRC-BMRF, Halifax, NS); Dr. Tara Sprules for 11.7 T NMR data acquisition for initial chemical shift assignment at the Québec Eastern Canada High Field NMR Facility (QANUC), supported by the Canada Foundation for Innovation (CFI), the Groupe de Recherche Axé sur la Structure des Protéines (GRASP), McGill University Faculty of Science and Department of Chemistry and PROTEO; and Dr. John Archibald for comments on the manuscript. This work was supported by Discovery and Research Tools and Instruments Grants from the Natural Sciences and Engineering Research Council of Canada (NSERC; to JKR, XQL, MP and MA); a Leaders Opportunity Fund award from the CFI (to JKR); a Dalhousie Medical Research Foundation Capital Equipment Grant (to JKR and XQL); research grants (to QM) from the National High Technology Research and Development Program 863 (NO 2006AA03Z451), the National Natural Science Foundation of China (NO 31070698), the Ph.D. Programs Foundation of Ministry of Education of China (No. 20120075110007); and, funds (to MP and MA) from PROTEO and CERMA. The TCI probes for the 16.4 T NMR spectrometer at the NRC-BMRF were provided by Dalhousie University through an Atlantic Canada Opportunities Agency Grant. JKR is supported by a Canadian Institutes for Health Research New Investigator Award; MLT by studentships from the Nova Scotia Health Research Foundation and an NSERC Postgraduate Scholarship; and, KEO by NSERC Undergraduate Student Research Awards.

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General

PubMed: MeSH publication types

Journal Article
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Cite this

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title = "Spider wrapping silk fibre architecture arising from its modular soluble protein precursor",

abstract = "Spiders store spidroins in their silk glands as high concentration aqueous solutions, spinning these dopes into fibres with outstanding mechanical properties. Aciniform (or wrapping) silk is the toughest spider silk and is devoid of the short amino acid sequence motifs characteristic of the other spidroins. Using solution-state NMR spectroscopy, we demonstrate that the 200 amino acid Argiope trifasciata AcSp1 repeat unit contrasts with previously characterized spidroins, adopting a globular 5-helix bundle flanked by intrinsically disordered N- and C-terminal tails. Split-intein-mediated segmental NMR-active isotope-enrichment allowed unambiguous demonstration of modular and malleable {"}beads-on-a-string{"} concatemeric behaviour. Concatemers form fibres upon manual drawing with silk-like morphology and mechanical properties, alongside secondary structuring and orientation consistent with native AcSp1 fibres. AcSp1 structural stability varies locally, with the fifth helix denaturing most readily. The structural transition of aciniform spidroin from a mostly α-helical dope to a mixed α-helix/β-sheet-containing fibre can be directly related to spidroin architecture and stability.",

author = "Tremblay, {Marie Laurence} and Lingling Xu and Thierry Lef{\`e}vre and Muzaddid Sarker and Orrell, {Kathleen E.} and J{\'e}r{\'e}mie Leclerc and Qing Meng and Michel P{\'e}zolet and Mich{\`e}le Auger and Liu, {Xiang Qin} and Rainey, {Jan K.}",

note = "Funding Information: Thanks to Drs. Stephen Bearne and David Waisman for CD spectropolarimeter access; Bruce Stewart for technical assistance; Xudong Dai for fibre mechanical properties testing; Dr. Mike Lumsden for 11.7 T NMR spectrometer support at the Nuclear Magnetic Resonance Research Resource Facility (NMR3, Dalhosie University); Ian Burton and Drs. Nadine Merkley and Ray Syvitski for 16.4 T NMR spectrometer support at the National Research Council Biological Magnetic Resonance Facility (NRC-BMRF, Halifax, NS); Dr. Tara Sprules for 11.7 T NMR data acquisition for initial chemical shift assignment at the Qu{\'e}bec Eastern Canada High Field NMR Facility (QANUC), supported by the Canada Foundation for Innovation (CFI), the Groupe de Recherche Ax{\'e} sur la Structure des Prot{\'e}ines (GRASP), McGill University Faculty of Science and Department of Chemistry and PROTEO; and Dr. John Archibald for comments on the manuscript. This work was supported by Discovery and Research Tools and Instruments Grants from the Natural Sciences and Engineering Research Council of Canada (NSERC; to JKR, XQL, MP and MA); a Leaders Opportunity Fund award from the CFI (to JKR); a Dalhousie Medical Research Foundation Capital Equipment Grant (to JKR and XQL); research grants (to QM) from the National High Technology Research and Development Program 863 (NO 2006AA03Z451), the National Natural Science Foundation of China (NO 31070698), the Ph.D. Programs Foundation of Ministry of Education of China (No. 20120075110007); and, funds (to MP and MA) from PROTEO and CERMA. The TCI probes for the 16.4 T NMR spectrometer at the NRC-BMRF were provided by Dalhousie University through an Atlantic Canada Opportunities Agency Grant. JKR is supported by a Canadian Institutes for Health Research New Investigator Award; MLT by studentships from the Nova Scotia Health Research Foundation and an NSERC Postgraduate Scholarship; and, KEO by NSERC Undergraduate Student Research Awards.",

year = "2015",

month = jun,

day = "26",

doi = "10.1038/srep11502",

language = "English",

volume = "5",

journal = "Scientific Reports",

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T1 - Spider wrapping silk fibre architecture arising from its modular soluble protein precursor

AU - Tremblay, Marie Laurence

AU - Xu, Lingling

AU - Lefèvre, Thierry

AU - Sarker, Muzaddid

AU - Orrell, Kathleen E.

AU - Leclerc, Jérémie

AU - Meng, Qing

AU - Pézolet, Michel

AU - Auger, Michèle

AU - Liu, Xiang Qin

AU - Rainey, Jan K.

N1 - Funding Information: Thanks to Drs. Stephen Bearne and David Waisman for CD spectropolarimeter access; Bruce Stewart for technical assistance; Xudong Dai for fibre mechanical properties testing; Dr. Mike Lumsden for 11.7 T NMR spectrometer support at the Nuclear Magnetic Resonance Research Resource Facility (NMR3, Dalhosie University); Ian Burton and Drs. Nadine Merkley and Ray Syvitski for 16.4 T NMR spectrometer support at the National Research Council Biological Magnetic Resonance Facility (NRC-BMRF, Halifax, NS); Dr. Tara Sprules for 11.7 T NMR data acquisition for initial chemical shift assignment at the Québec Eastern Canada High Field NMR Facility (QANUC), supported by the Canada Foundation for Innovation (CFI), the Groupe de Recherche Axé sur la Structure des Protéines (GRASP), McGill University Faculty of Science and Department of Chemistry and PROTEO; and Dr. John Archibald for comments on the manuscript. This work was supported by Discovery and Research Tools and Instruments Grants from the Natural Sciences and Engineering Research Council of Canada (NSERC; to JKR, XQL, MP and MA); a Leaders Opportunity Fund award from the CFI (to JKR); a Dalhousie Medical Research Foundation Capital Equipment Grant (to JKR and XQL); research grants (to QM) from the National High Technology Research and Development Program 863 (NO 2006AA03Z451), the National Natural Science Foundation of China (NO 31070698), the Ph.D. Programs Foundation of Ministry of Education of China (No. 20120075110007); and, funds (to MP and MA) from PROTEO and CERMA. The TCI probes for the 16.4 T NMR spectrometer at the NRC-BMRF were provided by Dalhousie University through an Atlantic Canada Opportunities Agency Grant. JKR is supported by a Canadian Institutes for Health Research New Investigator Award; MLT by studentships from the Nova Scotia Health Research Foundation and an NSERC Postgraduate Scholarship; and, KEO by NSERC Undergraduate Student Research Awards.

PY - 2015/6/26

Y1 - 2015/6/26

N2 - Spiders store spidroins in their silk glands as high concentration aqueous solutions, spinning these dopes into fibres with outstanding mechanical properties. Aciniform (or wrapping) silk is the toughest spider silk and is devoid of the short amino acid sequence motifs characteristic of the other spidroins. Using solution-state NMR spectroscopy, we demonstrate that the 200 amino acid Argiope trifasciata AcSp1 repeat unit contrasts with previously characterized spidroins, adopting a globular 5-helix bundle flanked by intrinsically disordered N- and C-terminal tails. Split-intein-mediated segmental NMR-active isotope-enrichment allowed unambiguous demonstration of modular and malleable "beads-on-a-string" concatemeric behaviour. Concatemers form fibres upon manual drawing with silk-like morphology and mechanical properties, alongside secondary structuring and orientation consistent with native AcSp1 fibres. AcSp1 structural stability varies locally, with the fifth helix denaturing most readily. The structural transition of aciniform spidroin from a mostly α-helical dope to a mixed α-helix/β-sheet-containing fibre can be directly related to spidroin architecture and stability.

AB - Spiders store spidroins in their silk glands as high concentration aqueous solutions, spinning these dopes into fibres with outstanding mechanical properties. Aciniform (or wrapping) silk is the toughest spider silk and is devoid of the short amino acid sequence motifs characteristic of the other spidroins. Using solution-state NMR spectroscopy, we demonstrate that the 200 amino acid Argiope trifasciata AcSp1 repeat unit contrasts with previously characterized spidroins, adopting a globular 5-helix bundle flanked by intrinsically disordered N- and C-terminal tails. Split-intein-mediated segmental NMR-active isotope-enrichment allowed unambiguous demonstration of modular and malleable "beads-on-a-string" concatemeric behaviour. Concatemers form fibres upon manual drawing with silk-like morphology and mechanical properties, alongside secondary structuring and orientation consistent with native AcSp1 fibres. AcSp1 structural stability varies locally, with the fifth helix denaturing most readily. The structural transition of aciniform spidroin from a mostly α-helical dope to a mixed α-helix/β-sheet-containing fibre can be directly related to spidroin architecture and stability.

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Spider wrapping silk fibre architecture arising from its modular soluble protein precursor

Abstract

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