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Title: Ambidextrous Helical Nanotubes from Self-Assembly of Designed Helical Hairpin Motifs

Abstract

Tandem repeat proteins exhibit native designability and represent potentially useful scaffolds for the construction of synthetic biomimetic assemblies. We have designed 2 synthetic peptides, HEAT_R1 and LRV_M3Δ1, based on the consensus sequences of single repeats of thermophilic HEAT (PBS_HEAT) and Leucine-Rich Variant (LRV) structural motifs, respectively. Self-assembly of the peptides afforded high-aspect ratio helical nanotubes. Cryo-electron microscopy with direct electron detection was employed to analyze the structures of the solvated filaments. The 3D reconstructions from the cryo-EM maps led to atomic models for the HEAT_R1 and LRV_M3Δ1 filaments at resolutions of 6.0 and 4.4 Å, respectively. Surprisingly, despite sequence similarity at the lateral packing interface, HEAT_R1 and LRV_M3Δ1 filaments adopt the opposite helical hand and differ significantly in helical geometry, while retaining a local conformation similar to previously characterized repeat proteins of the same class. The differences in the 2 filaments could be rationalized on the basis of differences in cohesive interactions at the lateral and axial interfaces. These structural data reinforce previous observations regarding the structural plasticity of helical protein assemblies and the need for high-resolution structural analysis. Despite these observations, the native designability of tandem repeat proteins offers the opportunity to engineer novel helical nanotubes. Moreover, the resultantmore » nanotubes have independently addressable and chemically distinguishable interior and exterior surfaces that would facilitate applications in selective recognition, transport, and release.« less

Authors:
 [1];  [2];  [3];  [1];  [3];  [3];  [3];  [4];  [3];  [1]
  1. Emory Univ., Atlanta, GA (United States)
  2. Brookhaven National Lab. (BNL), Upton, NY (United States)
  3. Univ. of Virginia, Charlottesville, VA (United States)
  4. Argonne National Lab. (ANL), Argonne, IL (United States)
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States); Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); National Science Foundation (NSF)
OSTI Identifier:
1525381
Alternate Identifier(s):
OSTI ID: 1563920
Report Number(s):
BNL-211762-2019-JAAM
Journal ID: ISSN 0027-8424
Grant/Contract Number:  
SC0012704; AC02-06CH11357
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 29; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Tandem Repeat Proteins; Cryo-EM; Helical Nanotube; Self-Assembly; cryo-EM; helical nanotube; self-assembly; tandem repeat proteins

Citation Formats

Hughes, Spencer, Wall, Joseph, Wang, Fengbin, Wang, Shengyuan, Kreutzberger, Mark, Osinski, Tomasz, Orlova, Albina, Zuo, Xiaobing, Egelman, Edward, and Conticello, Vincent. Ambidextrous Helical Nanotubes from Self-Assembly of Designed Helical Hairpin Motifs. United States: N. p., 2019. Web. doi:10.1073/pnas.1903910116.
Hughes, Spencer, Wall, Joseph, Wang, Fengbin, Wang, Shengyuan, Kreutzberger, Mark, Osinski, Tomasz, Orlova, Albina, Zuo, Xiaobing, Egelman, Edward, & Conticello, Vincent. Ambidextrous Helical Nanotubes from Self-Assembly of Designed Helical Hairpin Motifs. United States. https://doi.org/10.1073/pnas.1903910116
Hughes, Spencer, Wall, Joseph, Wang, Fengbin, Wang, Shengyuan, Kreutzberger, Mark, Osinski, Tomasz, Orlova, Albina, Zuo, Xiaobing, Egelman, Edward, and Conticello, Vincent. Mon . "Ambidextrous Helical Nanotubes from Self-Assembly of Designed Helical Hairpin Motifs". United States. https://doi.org/10.1073/pnas.1903910116. https://www.osti.gov/servlets/purl/1525381.
@article{osti_1525381,
title = {Ambidextrous Helical Nanotubes from Self-Assembly of Designed Helical Hairpin Motifs},
author = {Hughes, Spencer and Wall, Joseph and Wang, Fengbin and Wang, Shengyuan and Kreutzberger, Mark and Osinski, Tomasz and Orlova, Albina and Zuo, Xiaobing and Egelman, Edward and Conticello, Vincent},
abstractNote = {Tandem repeat proteins exhibit native designability and represent potentially useful scaffolds for the construction of synthetic biomimetic assemblies. We have designed 2 synthetic peptides, HEAT_R1 and LRV_M3Δ1, based on the consensus sequences of single repeats of thermophilic HEAT (PBS_HEAT) and Leucine-Rich Variant (LRV) structural motifs, respectively. Self-assembly of the peptides afforded high-aspect ratio helical nanotubes. Cryo-electron microscopy with direct electron detection was employed to analyze the structures of the solvated filaments. The 3D reconstructions from the cryo-EM maps led to atomic models for the HEAT_R1 and LRV_M3Δ1 filaments at resolutions of 6.0 and 4.4 Å, respectively. Surprisingly, despite sequence similarity at the lateral packing interface, HEAT_R1 and LRV_M3Δ1 filaments adopt the opposite helical hand and differ significantly in helical geometry, while retaining a local conformation similar to previously characterized repeat proteins of the same class. The differences in the 2 filaments could be rationalized on the basis of differences in cohesive interactions at the lateral and axial interfaces. These structural data reinforce previous observations regarding the structural plasticity of helical protein assemblies and the need for high-resolution structural analysis. Despite these observations, the native designability of tandem repeat proteins offers the opportunity to engineer novel helical nanotubes. Moreover, the resultant nanotubes have independently addressable and chemically distinguishable interior and exterior surfaces that would facilitate applications in selective recognition, transport, and release.},
doi = {10.1073/pnas.1903910116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 29,
volume = 116,
place = {United States},
year = {2019},
month = {7}
}

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