Amyloid-like fibrils from an α-helical transmembrane protein

Karen Stroobants; Janet Reiko Kumita; Nicola Harris; Dimitri Chirgadze; Christopher M Dobson; Paula Booth; Michele Vendruscolo

doi:10.1021/acs.biochem.7b00157

Amyloid-like fibrils from an α-helical transmembrane protein

Karen Stroobants, Janet Reiko Kumita, Nicola Harris, Dimitri Chirgadze, Christopher M Dobson, Paula Booth, Michele Vendruscolo

Chemistry

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17 Citations (Scopus)

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Abstract

The propensity to misfold and self-assemble into stable aggregates is increasingly recognised as a common feature of protein molecules. Our understanding of this phenomenon and of its links with human disease has increased substantially over the last two decades. Studies thus far, however, have been almost exclusively focussed on cytosolic proteins, resulting in a lack of detailed information about the misfolding and aggregation of membrane proteins. As a consequence, although such proteins make up approximately 30% of the human proteome and have high propensities to aggregate, relatively little is known about the biophysical nature of their assemblies. To shed light on this issue, we have studied as a model system an archetypical representative of the ubiquitous major facilitator superfamily, the Escherichia coli lactose permease (LacY). By using a combination of established indicators of cross-β structure and morphology, including the amyloid diagnostic dye thioflavin-T, circular dichroism, Fourier transform infrared spectroscopy, X-ray fibre diffraction, and transmission electron microscopy, we show that LacY can form amyloid-like fibrils under destabilising conditions. These results indicate that transmembrane α-helical proteins, as well as cytosolic proteins, have the ability to adopt this generic state.

Original language	English
Pages (from-to)	3225-3233
Journal	Biochemistry
Volume	56
Issue number	25
DOIs	https://doi.org/10.1021/acs.biochem.7b00157
Publication status	Published - 11 May 2017

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title = "Amyloid-like fibrils from an α-helical transmembrane protein",

abstract = "The propensity to misfold and self-assemble into stable aggregates is increasingly recognised as a common feature of protein molecules. Our understanding of this phenomenon and of its links with human disease has increased substantially over the last two decades. Studies thus far, however, have been almost exclusively focussed on cytosolic proteins, resulting in a lack of detailed information about the misfolding and aggregation of membrane proteins. As a consequence, although such proteins make up approximately 30% of the human proteome and have high propensities to aggregate, relatively little is known about the biophysical nature of their assemblies. To shed light on this issue, we have studied as a model system an archetypical representative of the ubiquitous major facilitator superfamily, the Escherichia coli lactose permease (LacY). By using a combination of established indicators of cross-β structure and morphology, including the amyloid diagnostic dye thioflavin-T, circular dichroism, Fourier transform infrared spectroscopy, X-ray fibre diffraction, and transmission electron microscopy, we show that LacY can form amyloid-like fibrils under destabilising conditions. These results indicate that transmembrane α-helical proteins, as well as cytosolic proteins, have the ability to adopt this generic state.",

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AU - Stroobants, Karen

AU - Kumita, Janet Reiko

AU - Harris, Nicola

AU - Chirgadze, Dimitri

AU - Dobson, Christopher M

AU - Booth, Paula

AU - Vendruscolo, Michele

PY - 2017/5/11

Y1 - 2017/5/11

N2 - The propensity to misfold and self-assemble into stable aggregates is increasingly recognised as a common feature of protein molecules. Our understanding of this phenomenon and of its links with human disease has increased substantially over the last two decades. Studies thus far, however, have been almost exclusively focussed on cytosolic proteins, resulting in a lack of detailed information about the misfolding and aggregation of membrane proteins. As a consequence, although such proteins make up approximately 30% of the human proteome and have high propensities to aggregate, relatively little is known about the biophysical nature of their assemblies. To shed light on this issue, we have studied as a model system an archetypical representative of the ubiquitous major facilitator superfamily, the Escherichia coli lactose permease (LacY). By using a combination of established indicators of cross-β structure and morphology, including the amyloid diagnostic dye thioflavin-T, circular dichroism, Fourier transform infrared spectroscopy, X-ray fibre diffraction, and transmission electron microscopy, we show that LacY can form amyloid-like fibrils under destabilising conditions. These results indicate that transmembrane α-helical proteins, as well as cytosolic proteins, have the ability to adopt this generic state.

AB - The propensity to misfold and self-assemble into stable aggregates is increasingly recognised as a common feature of protein molecules. Our understanding of this phenomenon and of its links with human disease has increased substantially over the last two decades. Studies thus far, however, have been almost exclusively focussed on cytosolic proteins, resulting in a lack of detailed information about the misfolding and aggregation of membrane proteins. As a consequence, although such proteins make up approximately 30% of the human proteome and have high propensities to aggregate, relatively little is known about the biophysical nature of their assemblies. To shed light on this issue, we have studied as a model system an archetypical representative of the ubiquitous major facilitator superfamily, the Escherichia coli lactose permease (LacY). By using a combination of established indicators of cross-β structure and morphology, including the amyloid diagnostic dye thioflavin-T, circular dichroism, Fourier transform infrared spectroscopy, X-ray fibre diffraction, and transmission electron microscopy, we show that LacY can form amyloid-like fibrils under destabilising conditions. These results indicate that transmembrane α-helical proteins, as well as cytosolic proteins, have the ability to adopt this generic state.

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DO - 10.1021/acs.biochem.7b00157

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EP - 3233

JO - Biochemistry

JF - Biochemistry

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ER -

Amyloid-like fibrils from an α-helical transmembrane protein

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