Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins

ABSTRACT Metal-reducing bacteria in the genus Geobacter use a complex protein apparatus to guide the self-assembly of a divergent type IVa pilin peptide and synthesize conductive pilus appendages that show promise for the sustainable manufacturing of protein nanowires. The preferential helical confo...

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Autores principales:	K. M. Cosert, Angelines Castro-Forero, Rebecca J. Steidl, Robert M. Worden, G. Reguera
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	type IV pili microbial nanowires nanotubes synthetic biology electromicrobiology Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/28200882ac7540928e248fe5493a6c1a
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spelling	oai:doaj.org-article:28200882ac7540928e248fe5493a6c1a2021-11-15T15:54:45ZBottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins10.1128/mBio.02721-192150-7511https://doaj.org/article/28200882ac7540928e248fe5493a6c1a2019-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02721-19https://doaj.org/toc/2150-7511ABSTRACT Metal-reducing bacteria in the genus Geobacter use a complex protein apparatus to guide the self-assembly of a divergent type IVa pilin peptide and synthesize conductive pilus appendages that show promise for the sustainable manufacturing of protein nanowires. The preferential helical conformation of the Geobacter pilin, its high hydrophobicity, and precise distribution of charged and aromatic amino acids are critical for biological self-assembly and conductivity. We applied this knowledge to synthesize via recombinant methods truncated pilin peptides for the bottom-up fabrication of protein nanowires and identified rate-limiting steps of pilin nucleation and fiber elongation that control assembly efficiency and nanowire length, respectively. The synthetic fibers retained the biochemical and electronic properties of the native pili even under chemical fixation, a critical consideration for integration of the nanowires into electronic devices. The implications of these results for the design and mass production of customized protein nanowires for diverse applications are discussed. IMPORTANCE The discovery in 2005 of conductive protein appendages (pili) in the metal-reducing bacterium Geobacter sulfurreducens challenged our understanding of biological electron transfer and pioneered studies in electromicrobiology that revealed the electronic basis of many microbial metabolisms and interactions. The protein nature of the pili afforded opportunities for engineering novel conductive peptides for the synthesis of nanowires via cost-effective and scalable manufacturing approaches. However, methods did not exist for efficient production, purification, and in vitro assembly of pilins into nanowires. Here we describe platforms for high-yield recombinant synthesis of Geobacter pilin derivatives and their assembly as protein nanowires with biochemical and electronic properties rivaling those of the native pili. The bottom-up fabrication of protein nanowires exclusively from pilin building blocks confirms unequivocally the charge transport capacity of the peptide assembly and establishes the intellectual foundation needed to manufacture pilin-based nanowires in bioelectronics and other applications.K. M. CosertAngelines Castro-ForeroRebecca J. SteidlRobert M. WordenG. RegueraAmerican Society for Microbiologyarticletype IV pilimicrobial nanowiresnanotubessynthetic biologyelectromicrobiologyMicrobiologyQR1-502ENmBio, Vol 10, Iss 6 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	type IV pili microbial nanowires nanotubes synthetic biology electromicrobiology Microbiology QR1-502
spellingShingle	type IV pili microbial nanowires nanotubes synthetic biology electromicrobiology Microbiology QR1-502 K. M. Cosert Angelines Castro-Forero Rebecca J. Steidl Robert M. Worden G. Reguera Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins
description	ABSTRACT Metal-reducing bacteria in the genus Geobacter use a complex protein apparatus to guide the self-assembly of a divergent type IVa pilin peptide and synthesize conductive pilus appendages that show promise for the sustainable manufacturing of protein nanowires. The preferential helical conformation of the Geobacter pilin, its high hydrophobicity, and precise distribution of charged and aromatic amino acids are critical for biological self-assembly and conductivity. We applied this knowledge to synthesize via recombinant methods truncated pilin peptides for the bottom-up fabrication of protein nanowires and identified rate-limiting steps of pilin nucleation and fiber elongation that control assembly efficiency and nanowire length, respectively. The synthetic fibers retained the biochemical and electronic properties of the native pili even under chemical fixation, a critical consideration for integration of the nanowires into electronic devices. The implications of these results for the design and mass production of customized protein nanowires for diverse applications are discussed. IMPORTANCE The discovery in 2005 of conductive protein appendages (pili) in the metal-reducing bacterium Geobacter sulfurreducens challenged our understanding of biological electron transfer and pioneered studies in electromicrobiology that revealed the electronic basis of many microbial metabolisms and interactions. The protein nature of the pili afforded opportunities for engineering novel conductive peptides for the synthesis of nanowires via cost-effective and scalable manufacturing approaches. However, methods did not exist for efficient production, purification, and in vitro assembly of pilins into nanowires. Here we describe platforms for high-yield recombinant synthesis of Geobacter pilin derivatives and their assembly as protein nanowires with biochemical and electronic properties rivaling those of the native pili. The bottom-up fabrication of protein nanowires exclusively from pilin building blocks confirms unequivocally the charge transport capacity of the peptide assembly and establishes the intellectual foundation needed to manufacture pilin-based nanowires in bioelectronics and other applications.
format	article
author	K. M. Cosert Angelines Castro-Forero Rebecca J. Steidl Robert M. Worden G. Reguera
author_facet	K. M. Cosert Angelines Castro-Forero Rebecca J. Steidl Robert M. Worden G. Reguera
author_sort	K. M. Cosert
title	Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins
title_short	Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins
title_full	Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins
title_fullStr	Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins
title_full_unstemmed	Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins
title_sort	bottom-up fabrication of protein nanowires via controlled self-assembly of recombinant <italic toggle="yes">geobacter</italic> pilins
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/28200882ac7540928e248fe5493a6c1a
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Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant <italic toggle="yes">Geobacter</italic> Pilins

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