Assembly of transmembrane pores from mirror-image peptides

Smrithi Krishnan R, Kalyanashis Jana, Amina H. Shaji, Karthika S. Nair, Anjali Devi Das, Devika Vikraman, Harsha Bajaj, Ulrich Kleinekathöfer and Kozhinjampara R. Mahendran ()
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Smrithi Krishnan R: Rajiv Gandhi Centre for Biotechnology
Kalyanashis Jana: Jacobs University Bremen
Amina H. Shaji: Rajiv Gandhi Centre for Biotechnology
Karthika S. Nair: CSIR- National Institute for Interdisciplinary Science and Technology (NIIST)
Anjali Devi Das: Rajiv Gandhi Centre for Biotechnology
Devika Vikraman: Rajiv Gandhi Centre for Biotechnology
Harsha Bajaj: CSIR- National Institute for Interdisciplinary Science and Technology (NIIST)
Ulrich Kleinekathöfer: Jacobs University Bremen
Kozhinjampara R. Mahendran: Rajiv Gandhi Centre for Biotechnology

Nature Communications, 2022, vol. 13, issue 1, 1-13

Abstract: Abstract Tailored transmembrane alpha-helical pores with desired structural and functional versatility have promising applications in nanobiotechnology. Herein, we present a transmembrane pore DpPorA, based on the natural pore PorACj, built from D-amino acid α-helical peptides. Using single-channel current recordings, we show that DpPorA peptides self-assemble into uniform cation-selective pores in lipid membranes and exhibit properties distinct from their L-amino acid counterparts. DpPorA shows resistance to protease and acts as a functional nanopore sensor to detect cyclic sugars, polypeptides, and polymers. Fluorescence imaging reveals that DpPorA forms well-defined pores in giant unilamellar vesicles facilitating the transport of hydrophilic molecules. A second D-amino acid peptide based on the polysaccharide transporter Wza forms transient pores confirming sequence specificity in stable, functional pore formation. Finally, molecular dynamics simulations reveal the specific alpha-helical packing and surface charge conformation of the D-pores consistent with experimental observations. Our findings will aid the design of sophisticated pores for single-molecule sensing related technologies.

Date: 2022
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DOI: 10.1038/s41467-022-33155-6

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