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1.
Methods Mol Biol ; 2430: 3-16, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35476322

RESUMO

Filamentous microtubules, polymers of the heterodimeric protein tubulins play one of the major roles in the emergent nano-biotechnological devices. To develop the feature of those devices, it is important to understand the function of microtubule in in vitro, hence, the availability of purified αß-tubulin is required. Additionally, fluorescently labeled tubulin has become a powerful approach for extensively studying the dynamics of these components. In this chapter, the process of purifying the heterodimeric αß-tubulin from porcine brain will be described, as well as the process of labeling of the purified tubulin with fluorescence dye.


Assuntos
Corantes Fluorescentes , Tubulina (Proteína) , Animais , Encéfalo/metabolismo , Fluorescência , Corantes Fluorescentes/metabolismo , Microtúbulos/metabolismo , Suínos , Tubulina (Proteína)/metabolismo
2.
Methods Mol Biol ; 2430: 47-59, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35476324

RESUMO

The filamentous cytoskeletal protein microtubule, a polymer of α and ß heterodimers of tubulin, plays major roles in intracellular transport as well as in vitro molecular actuation and transportation. Functionalization of tubulin dimers through covalent linkage facilitates utilization of microtubule in the nanobioengineering. Here we present a detailed description of the methodologies used to modify tubulin dimers with DNA strand and biotin through covalent interaction.


Assuntos
Biotina , Tubulina (Proteína) , Transporte Biológico , Biotina/metabolismo , DNA/metabolismo , Microtúbulos/metabolismo , Tubulina (Proteína)/metabolismo
3.
Methods Mol Biol ; 2430: 231-240, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35476336

RESUMO

Fabrication of molecular devices using biomolecules through biomimetic approaches has witnessed a surge in interest in recent years. DNA a versatile programmable material offers an opportunity to realize complicated operations through the designing of various nanostructures such as DNA origami. Here we describe the methods to use DNA origami for the self-assembly of the biomolecular motor system, microtubule (MT)-kinesin. A rodlike DNA origami motif facilitates the self-assembly of MTs into asters. A smooth muscle like molecular contraction system could be realized following the method where DNA mediated self-assembly of MTs permits dynamic contraction in the presence of kinesins through an energy dissipative process.


Assuntos
Cinesinas , Nanoestruturas , DNA/química , Microtúbulos/química , Músculo Liso , Nanoestruturas/química
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