Engineering tubulin: microtubule functionalization approaches for nanoscale device applications
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With the emergences of engineered devices at microscale and nanoscale dimensions, there is a growing need for controlled actuation and transport at these length scales. The kinesin–microtubule system provides a highly evolved biological transport system well suited for these tasks. Accordingly, there is an ongoing effort to create hybrid nanodevices that integrate biological components with engineered materials for applications such as biological separations, nanoscale assembly, and sensing. Adopting microtubules for these applications generally requires covalent attachment of biotin, fluorophores, or other biomolecules to tubulin enable surface or cargo attachment, or visualization. This review summarizes different strategies for functionalizing microtubules for application-focused as well as basic biological research. These functionalization strategies must maintain the integrity of microtubule proteins so that they do not depolymerize and can be transported by kinesin motors, while adding utility such as the ability to reversibly bind cargo. The relevant biochemical and electrical properties of microtubules are discussed, as well as strategies for microtubule stabilization and long-term storage. Next, attachment strategies, such as antibodies and DNA hybridization that have proven useful to date, are discussed in the context of ongoing hybrid nanodevice research. The review concludes with a discussion of less explored opportunities, such as harnessing the utility of tubulin posttranslational modifications and the use of recombinant tubulin that may enable future progress in nanodevice development.
KeywordsFluorescence Biotinylation Microfabrication Nanotechnology Kinesin Cytoskeleton
J.L.M was supported by NSF grant MCB 0920911 and W.O.H was supported by NIH grant GM083297.
- Agarwal A, Hess H (2010b) Molecular motors as components of future medical devices and engineered materials. J Nanotech Eng Med 1:1–9Google Scholar
- Hess H, Vogel V (2001) Molecular shuttles based on motor proteins: active transport in synthetic environments. J Biotechnol 82:67–85Google Scholar
- Hiller Y, Gershoni JM, Bayer EA, Wilchek M (1987) Biotin binding to avidin. Oligosaccharide side chain not required for ligand association. Biochem J 248:167–171Google Scholar
- Kellogg DR, Mitchison TJ, Alberts BM (1988) Behaviour of microtubules and actin filaments in living Drosophila embryos. Development 103:675–686Google Scholar
- Panchuk-Voloshina N, Haugland RP, Bishop-Stewart J, Bhalgat MK, Millard PJ, Mao F, Leung WY (1999) Alexa dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates. J Histochem Cytochem 47:1179–1188Google Scholar
- Uppalapati M, Huang YM, Shastry S, Jackson TN, Hancock WO (2009) In: Zahn JD, Lee LP (eds) Microtubule motors in microfluidics. Methods in bioengineering: microfabrication and microfluidics. Artech House, BostonGoogle Scholar
- Yokokawa R, Takeuchi S, Kon T, Ohkura R, Edamatsu M, Sutoh K and Fujita H (2003) Transportation of micromachined structures by biomolecular linear motors. Proceedings Micro Electro Mechanical Systems, pp 8–11Google Scholar