Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Accelerating the design of biomimetic materials by integrating RNA-seq with proteomics and materials science

Abstract

Efforts to engineer new materials inspired by biological structures are hampered by the lack of genomic data from many model organisms studied in biomimetic research. Here we show that biomimetic engineering can be accelerated by integrating high-throughput RNA-seq with proteomics and advanced materials characterization. This approach can be applied to a broad range of systems, as we illustrate by investigating diverse high-performance biological materials involved in embryo protection, adhesion and predation. In one example, we rapidly engineer recombinant squid sucker ring teeth proteins into a range of structural and functional materials, including nanopatterned surfaces and photo-cross-linked films that exceed the mechanical properties of most natural and synthetic polymers. Integrating RNA-seq with proteomics and materials science facilitates the molecular characterization of natural materials and the effective translation of their molecular designs into a wide range of bio-inspired materials.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Integrating RNA-seq, proteomics and material science.
Figure 2: Marine snail egg-case bioelastomeric membranes and proteins.
Figure 3: Identification of green mussel (P. viridis) adhesive protein sequences.
Figure 4: Sucker ring teeth biomaterials from jumbo squid.
Figure 5: Solvent-free processing of thermoplastic suckerin.
Figure 6: Engineering of recombinant suckerin-39.

Similar content being viewed by others

Accession codes

Accessions

Protein Data Bank

References

  1. Huebsch, N. & Mooney, D.J. Inspiration and application in the evolution of biomaterials. Nature 462, 426–432 (2009).

    Article  CAS  Google Scholar 

  2. Dunlop, J., Weinkamer, R. & Fratzl, P. Artful interfaces within biological materials. Mater. Today 14, 70–78 (2010).

    Article  Google Scholar 

  3. Cranford, S.W., De Boer, J., Van Blitterswijk, C. & Buehler, M.J. Materiomics: an -omics approach to biomaterials research. Adv. Mater. 25, 802–824 (2013).

    Article  CAS  Google Scholar 

  4. Boyle, A. & Woolfson, D.N. De novo designed peptides for biological applications. Chem. Soc. Rev. 40, 4295–4306 (2011).

    Article  CAS  Google Scholar 

  5. Guvendiren, M., Messersmith, P.B. & Shull, K.R. Self-assembly and adhesion of DOPA-modified methacrylic triblock hydrogels. Biomacromolecules 9, 122–128 (2008).

    Article  CAS  Google Scholar 

  6. Hildebrand, M. Diatoms, biomineralization processes, and genomics. Chem. Rev. 108, 4855–4874 (2008).

    Article  CAS  Google Scholar 

  7. Suzuki, M. et al. An acidic matrix protein, pif, is a key macromolecule for nacre formation. Science 325, 1388–1390 (2009).

    Article  CAS  Google Scholar 

  8. Lee, B.P., Messersmith, P.B., Israelachvili, J.N. & Waite, J.H. Mussel-inspired adhesives and coatings. Annu. Rev. Mater. Res. 41, 99–132 (2011).

    Article  CAS  Google Scholar 

  9. Waite, J.H. & Broomell, C.C. Changing environments and structure–property relationships in marine biomaterials. J. Exp. Biol. 215, 873–883 (2012).

    Article  CAS  Google Scholar 

  10. Metzker, M.L. Sequencing technologies—the next generation. Nat. Rev. Genet. 11, 31–46 (2010).

    Article  CAS  Google Scholar 

  11. Ozsolak, F. & Milos, P.M. RNA sequencing: advances, challenges and opportunities. Nat. Rev. Genet. 12, 87–98 (2011).

    Article  CAS  Google Scholar 

  12. Wang, Z., Gerstein, M. & Snyder, M. RNA-seq: a revolutionary tool for transcriptomics. Nat. Rev. Genet. 10, 57–63 (2009).

    Article  CAS  Google Scholar 

  13. Garber, M., Grabherr, M.G., Guttman, M. & Trapnell, C. Computational methods for transcriptome annotation and quantification using RNA-seq. Nat. Methods 8, 469–477 (2011).

    Article  CAS  Google Scholar 

  14. Miserez, A., Wasko, S., Carpenter, C. & Waite, J.H. Non-entropic and reversible long-range deformation of an encapsulating bioelastomer. Nat. Mater. 8, 910–916 (2009).

    Article  CAS  Google Scholar 

  15. Miserez, A. & Guerette, P.A. Phase transition-induced elasticity of α-helical bioelastomeric fibres and networks. Chem. Soc. Rev. 42, 1973–1995 (2013).

    Article  CAS  Google Scholar 

  16. Scotto-Lavino, E., Du, G. & Frohman, M.A. 3′ End cDNA amplification using classic RACE. Nat. Protoc. 1, 2742–2745 (2006).

    Article  CAS  Google Scholar 

  17. Armstrong, C.T., Vincent, T.L., Green, P.J. & Woolfson, D.N. SCORER 2.0: an agorithm for distinguishing parallel dimeric and trimeric coiled-coil sequences. Bioinformatics 27, 1908–1914 (2011).

    Article  CAS  Google Scholar 

  18. Arslan, M., Qin, Z. & Buehler, M.J. Coiled-coil intermediate filament stutter instability and molecular unfolding. Comput. Methods Biomech. Biomed. Engin. 14, 483–489 (2011).

    Article  Google Scholar 

  19. Waite, J.H., Holten-Andersen, N., Jewhurst, S.A. & Sun, C. Mussel adhesion: finding the tricks worth mimicking. J. Adhes. 81, 297–317 (2005).

    Article  CAS  Google Scholar 

  20. Harrington, M.J. & Waite, J.H. Holdfast heroics: comparing the molecular and mechanical properties of Mytilus californianus byssal threads. J. Exp. Biol. 210, 4307–4318 (2007).

    Article  CAS  Google Scholar 

  21. Zhao, H., Robertson, N., Jewhurst, S.A. & Waite, J.H. Probing the adhesive footprints of Mytilus californianus byssus. J. Biol. Chem. 281, 11090–11096 (2006).

    Article  CAS  Google Scholar 

  22. Zhao, H. & Waite, J.H. Proteins in load-bearing junctions: the histidine-rich metal-binding protein of mussel byssus. Biochemistry 45, 14223–14231 (2006).

    Article  CAS  Google Scholar 

  23. Yu, J. et al. Mussel protein adhesion depends on interprotein thiol-mediated redox modulation. Nat. Chem. Biol. 7, 588–590 (2011).

    Article  CAS  Google Scholar 

  24. Miserez, A. et al. Microstructural and biochemical characterization of the nano-porous sucker rings from Dosidicus gigas. Adv. Mater. 21, 401–406 (2009).

    Article  CAS  Google Scholar 

  25. Guerette, P., Gizinger, D., Weber, B. & Gosline, J.M. Silk properties determined by gland-specific expression of a spider fibroin gene family. Science 272, 112–115 (1996).

    Article  CAS  Google Scholar 

  26. Gatesy, J., Hayashi, C.Y., Motriuk, D., Woods, J. & Lewis, R. Extreme diversity, conservation, and convergence of spider silk fibroin sequences. Science 291, 2603–2605 (2001).

    Article  CAS  Google Scholar 

  27. Yano, M., Nagai, K., Morimoto, K. & Miyamoto, K. Shematrin: a family of glycine-rich structural proteins in the shell of the pearl oyster Pinctada fucata. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 144, 254–262 (2006).

    Article  Google Scholar 

  28. Keten, S., Xu, Z., Ihle, B. & Buehler, M.J. Nanoconfinement controls stiffness, strength and mechanical toughness of beta-sheet crystals in silk. Nat. Mater. 9, 359–367 (2010).

    Article  CAS  Google Scholar 

  29. Rousseau, M.-E., Lefevre, T., Beaulieu, L., Asakura, T. & Pezolet, M. Study of protein conformation and orientation in silkworm and spider silk fibers using raman microspectroscopy. Biomacromolecules 5, 2247–2257 (2004).

    Article  CAS  Google Scholar 

  30. Steinhart, M. et al. Polymer nanotubes by wetting of ordered porous templates. Science 296, 1997 (2002).

    Article  CAS  Google Scholar 

  31. Omenetto, F.G. & Kaplan, D. New opportunities for an ancient material. Science 329, 528–531 (2010).

    Article  CAS  Google Scholar 

  32. Tao, H., Kaplan, D.L. & Omenetto, F.G. Silk materials–a road to sustainable high technology. Adv. Mater. 24, 2824–2837 (2012).

    Article  CAS  Google Scholar 

  33. Gosline, J.M., Guerette, P.A., Ortlepp, C.S. & Savage, K.N. The mechanical design of spider silks: from fibroins sequence to mechanical function. J. Exp. Biol. 202, 3295–3303 (1999).

    CAS  PubMed  Google Scholar 

  34. Fancy, D.A. & Kodadek, T. Chemistry for the analysis of protein-protein interactions: rapid and efficient cross-linking triggered by long wavelength light. Proc. Natl. Acad. Sci. USA 96, 6020–6024 (1999).

    Article  CAS  Google Scholar 

  35. Savage, K.N. & Gosline, J.M. The effect of proline on the network structure of major ampullate silks as inferred from their mechanical and optical properties. J. Exp. Biol. 211, 1937–1947 (2008).

    Article  CAS  Google Scholar 

  36. Zok, F. & Miserez, A. Property maps for abrasion resistance of materials. Acta Mater. 55, 6365–6371 (2007).

    Article  CAS  Google Scholar 

  37. Hancock, M., Sekeroglu, K. & Demirel, M.C. Bioinspired directional surfaces for adhesion, wetting and transport. Adv. Funct. Mater. 22, 2223–2234 (2012).

    Article  CAS  Google Scholar 

  38. Engler, A.J.S.S., Sweeney, H.L. & Discher, D.E. Matrix elasticity directs stem cell lineage specification. Cell 126, 677–689 (2006).

    Article  CAS  Google Scholar 

  39. Kim, S.J. et al. Silk inverse opals. Nat. Photonics 6, 818–823 (2012).

    Article  CAS  Google Scholar 

  40. Zhang, J. et al. Stabilization of vaccines and antibiotics in silk and eliminating the cold chain. Proc. Natl. Acad. Sci. USA 109, 11981–11986 (2012).

    Article  CAS  Google Scholar 

  41. Dickerson, M.B., Sandhage, K.H. & Naik, R.R. Protein- and peptide-directed syntheses of inorganic materials. Chem. Rev. 108, 4935–4978 (2008).

    Article  CAS  Google Scholar 

  42. Grabherr, M.G. et al. Full-length transcriptome assembly from RNA-seq data without a reference genome. Nat. Biotechnol. 29, 644–652 (2011).

    Article  CAS  Google Scholar 

  43. Li, B. & Dewey, C.N. RSEM: accurate transcript quantification from RNA-seq data with or without a reference genome. BMC Bioinformatics 12, 323 (2011).

    Article  CAS  Google Scholar 

  44. Edgar, R.C. Search and custering orders of magnitude faster than BLAST. Bioinformatics 26, 2460–2461 (2010).

    Article  CAS  Google Scholar 

  45. Ma, B. et al. PEAKS: powerful software for peptide de novo sequencing by MS/MS. Rapid Commun. Mass Spectrom. 17, 2337–2342 (2003).

    Article  CAS  Google Scholar 

  46. Hwang, D.S. et al. Protein- and metal-dependent interactions of a prominent protein in mussel adhesive plaques. J. Biol. Chem. 285, 25850–25858 (2010).

    Article  CAS  Google Scholar 

  47. Waite, J.H. & Tanzer, M.L. Specific colorimetric detection of o-diphenols and DOPA-containing peptides. Anal. Biochem. 111, 131–136 (1981).

    Article  CAS  Google Scholar 

  48. Larkin, M.A. et al. Clustal W and Clustal X version 2.0. Bioinformatics 23, 2947–2948 (2007).

    Article  CAS  Google Scholar 

  49. Roy, A., Kucukural, A. & Zhang, Y. I-TASSER: a unified platform for automated protein structure and function prediction. Nat. Protoc. 5, 725–738 (2010).

    Article  CAS  Google Scholar 

  50. Jaenicke, E., Buchler, K., Markl, J., Decker, H. & Barends, T.R.M. Cupredoxin-like domains in haemocyanins. Biochem. J. 426, 373–378 (2010).

    Article  CAS  Google Scholar 

  51. Matoba, Y. et al. A molecular mechanism for copper transportation to tyrosinase that is assisted by a metallochaperone, caddie protein. J. Biol. Chem. 286, 30219–30231 (2011).

    Article  CAS  Google Scholar 

  52. Sendovski, M., Kanteev, M., Shuster Ben-Yosef, V., Adir, N. & Fishman, A. First structures of an active bacterial tyrosinase reveal copper plasticity. J. Mol. Biol. 405, 227–237 (2011).

    Article  CAS  Google Scholar 

  53. Cuff, M.E., Miller, K.I., van Holde, K.E. & Hendrickson, W.A. Crystal structure of a functional unit from octopus hemocyanin. J. Mol. Biol. 278, 855–870 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to H. Waite and S. Brenner for insightful discussions and suggestions. We thank J. Weaver, C. Salinas and D. DeMartini for help with Jumbo squid sucker teeth sample collection. G. Muniraj, L. Sellou and C. Sutanto assisted with extraction and purification of P. viridis and squid sucker teeth proteins. V. Gangu provided informatics support and S. Seah provided assistance with tyrosinase modeling. V. Lipik assisted with the acquisition of MALDI-TOF data. W.M. Guo helped with cell culture experiments. We thank K.S. Tan for his advice on snail biology and animal collection. V. Seow, E. Tay and A. Hermawan helped with sample collection and mechanical testing of P. cochlidium egg cases. This research was funded by the Singapore National Research Foundation (NRF) through a NRF Fellowship (A. Miserez), a Singapore Ministry of Education Tier 2 grant (A. Miserez, MOE2011-T2-2-044) and the Agency for Science, Technology and Research (A*Star) of Singapore (S.H., Y.S., F.T.W., V.H.B.H.). M.C.D. and A.P.-F. were supported by US National Institutes of Health grant no. 1R21HL112114-01.

Author information

Authors and Affiliations

Authors

Contributions

P.A.G. and S.H. conducted molecular experiments and analyzed sequencing data. S.H. and K.W.K. generated and compiled RNA-seq data. A. Miserez and S.A. conducted and analyzed mechanical testing experiments. Y.S. conducted tyrosinase assays, conducted and analyzed qPCR data, and provided editorial comments. M.R. performed LC MS/MS experiments and analyzed the data with PEAKS. A. Masic conducted and interpreted Raman spectroscopy data. M.C.D. developed the idea of microwave heating of SRT proteins. A.P.-F. and M.C.D. conducted and analyzed the template wetting experiments with native and recombinant proteins and provided editorial comments. P.A.G. conceived and conducted the suckerin fiber and film engineering and characterization experiments and developed and conducted the photo cross-linking experiments. S.A. and G.Z.T. conducted Raman and FTIR experiments on drawn fibers. F.T.W. performed structural modeling of tyrosinases. K.W.K. and D.D. expressed and purified recombinant proteins. V.H.B.H. helped perform and analyze cell culture experiments. A. Miserez, P.A.G. and S.H. designed the study and wrote the paper.

Corresponding author

Correspondence to Ali Miserez.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 and Supplementary Tables 1 and 2 (PDF 7181 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guerette, P., Hoon, S., Seow, Y. et al. Accelerating the design of biomimetic materials by integrating RNA-seq with proteomics and materials science. Nat Biotechnol 31, 908–915 (2013). https://doi.org/10.1038/nbt.2671

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt.2671

This article is cited by

Search

Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research