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Split-Intein Triggered Protein Hydrogels

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Book cover Split Inteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1495))

Abstract

Proteins are nature’s building blocks and indispensable in living organisms. Protein-based hydrogels have a wide variety of applications in research and biotechnology. In this chapter, we describe an intein-mediated protein hydrogel that utilizes two synthetic soluble protein block copolymers, each containing a subunit of a trimeric protein that serves as a cross-linker and one half of the naturally split DnaE intein from Nostoc punctiforme. Mixing of these two protein block copolymers initiates an intein trans-splicing reaction that constitutes a self-assembling polypeptide flanked by cross-linkers, triggering protein hydrogel formation. The generated hydrogels are highly stable under both acidic and basic conditions, and at temperatures up to 50 °C. In addition, these hydrogels are able to undergo rapid reassembly after shear-induced rupture. Incorporation of an appropriate binding motif into the protein block copolymers enables the convenient site-specific incorporation of functional globular proteins into the hydrogel network.

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Acknowledgements

This work was supported in part by the National Science Foundation CAREER award, US Air force YIP and Norman Hackman Advanced Research Program. We would like to acknowledge Dr. David Tirrell (Caltech) for his kind gift of the plasmid pQE9 AC10Atrp [16], Dr. Takehisa Matsuda (Kanazawa Institute of Technology, Hakusan, Ishikawa, Japan) for his kind gift of the plasmid pET30-CutA-Tip1 [19], Dr. Tom Muir (Princeton University) for his kind gift of the plasmid KanR-IntRBS-NpuNC-CFN [20], and Dr. Jay D. Keasling (UC Berkley) for his kind gift of the plasmid pJD757 [21].

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Authors and Affiliations

  1. Department of Chemical Engineering, Texas A&M University, College Station, TX, 77843, USA

    Miguel A. Ramirez & Zhilei Chen

  2. Department of Microbial and Molecular Pathogenesis, Texas A&M University Health Science Center, College Station, TX, 77843, USA

    Zhilei Chen

Authors
  1. Miguel A. Ramirez
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  2. Zhilei Chen
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Corresponding author

Correspondence to Zhilei Chen .

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Editors and Affiliations

  1. Institute of Biochemistry Department of Chemistry and Pharmacy, University of Muenster, Münster, Germany

    Henning D. Mootz

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Ramirez, M.A., Chen, Z. (2017). Split-Intein Triggered Protein Hydrogels. In: Mootz, H. (eds) Split Inteins. Methods in Molecular Biology, vol 1495. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6451-2_11

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  • DOI: https://doi.org/10.1007/978-1-4939-6451-2_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6449-9

  • Online ISBN: 978-1-4939-6451-2

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