Labile assembly of a tardigrade protein induces biostasis

Sánchez-Martínez, Silvia; Nguyen, Kenny H.; Biswas, Sourav; Nicholson, Vincent; Romanyuk, Andrey; Ramirez, John F; Kc, Shraddha; Akter, Afroza; Childs, Charles A.; Usher, Grace A.; Ginell, Garrett M.; Yu, Feng; Gollub, Edith G.; Malferrari, Marco; Francia, Francesco; Venturoli, Giovanni; Martin, Erik W.; Caporaletti, Federico; Giubertoni, Giulia; Woutersen, Sander; Sukenik, Shahar; Woolfson, Derek N.; Holehouse, Alex S.; Boothby, Thomas E.

doi:10.1101/2023.06.30.547219

Cited by 4 publications

(28 citation statements)

References 104 publications

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“…Biophysical and computational studies have shown that CAHS D occupies a dumbbell‐like ensemble of interconverting conformations composed of two relatively compact termini bridged by an internal linker region (Figure 1a) (Eicher et al, 2022; Malki et al, 2022; Sanchez‐Martinez, Nguyen, et al, 2023). The terminal regions of CAHS D contain transient helical and beta‐structure, while the linker region interconverts between random coil and helical structures (Eicher et al, 2022; Malki et al, 2022; Sanchez‐Martinez, Nguyen, et al, 2023) (Figure 1a). The CAHS linker contains both polar and hydrophobic residues, and when in a helical state, this region exists as an amphipathic helix, as quantified by the hydrophobic moment (Figure 1a).…”

Section: Resultsmentioning

confidence: 99%

“…CAHS D, a model CAHS protein. (a) Representative conformational ensemble of model CAHS D protein by all‐atom Monte Carlo simulation (Sanchez‐Martinez, Nguyen, et al, 2023). Helix/sheet assignments made using JPRED4 (Drozdetskiy et al, 2015).…”

Section: Resultsmentioning

confidence: 99%

“…Since CAHS D (as well as other CAHS proteins) is known to be partially helical (Eicher et al, 2022; Malki et al, 2022; Sanchez‐Martinez, Nguyen, et al, 2023; Tanaka et al, 2022; Yamaguchi et al, 2012) and helicity has been proposed to be linked to the protective capacity of IDPs during drying (LeBlanc & Hand, 2021), we asked if the helicity of our variants correlates with protection. To test this, we first performed circular dichroism (CD) spectroscopy on each of our variants (Figure 3b) (File S5).…”

Section: Resultsmentioning

confidence: 99%

“…Amphipathicity of LEA proteins has previously been proposed to allow for hydrophobic interactions between LEA proteins and client molecules (e.g., membrane headgroups) and could play a role in self‐assembly of proteins into oligomers (Shou et al, 2019; Tunnacliffe & Wise, 2007). While no evidence to date exists that CAHS D, or its linker region, interacts with membranes, several studies have demonstrated that CAHS D, as well as other CAHS proteins, oligomerize (Eicher et al, 2022; Malki et al, 2022; Sanchez‐Martinez, Nguyen, et al, 2023; Tanaka et al, 2022; Yagi‐Utsumi et al, 2021). Furthermore, the helicity and amphipathic nature of the linker region of CAHS D and other CAHS proteins has been shown to be essential for oligomerization into higher‐order structures (Sanchez‐Martinez, Nguyen, et al, 2023; Tanaka et al, 2022).…”

Section: Discussionmentioning

confidence: 99%

“…While no evidence to date exists that CAHS D, or its linker region, interacts with membranes, several studies have demonstrated that CAHS D, as well as other CAHS proteins, oligomerize (Eicher et al, 2022; Malki et al, 2022; Sanchez‐Martinez, Nguyen, et al, 2023; Tanaka et al, 2022; Yagi‐Utsumi et al, 2021). Furthermore, the helicity and amphipathic nature of the linker region of CAHS D and other CAHS proteins has been shown to be essential for oligomerization into higher‐order structures (Sanchez‐Martinez, Nguyen, et al, 2023; Tanaka et al, 2022). Thus, one might speculate that the reason behind a decrease in protection upon helix‐disruption is due to a loss in the ability to oligomerize.…”

Section: Discussionmentioning

confidence: 99%

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Helicity of a tardigrade disordered protein contributes to its protective function during desiccation

Biswas,

Gollub,

et al. 2024

Protein Science

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In order to survive extreme drying (anhydrobiosis), many organisms, spanning every kingdom of life, accumulate intrinsically disordered proteins (IDPs). For decades, the ability of anhydrobiosis‐related IDPs to form transient amphipathic helices has been suggested to be important for promoting desiccation tolerance. However, evidence empirically supporting the necessity and/or sufficiency of helicity in mediating anhydrobiosis is lacking. Here we demonstrate that the linker region of CAHS D, a desiccation‐related IDP from the tardigrade Hypsibius exemplaris, that contains significant helical structure, is the protective portion of this protein. Perturbing the sequence composition and grammar of the linker region of CAHS D, through the insertion of helix‐breaking prolines, modulating the identity of charged residues, or replacement of hydrophobic amino acids with serine or glycine residues results in variants with different degrees of helical structure. Importantly, correlation of protective capacity and helical content in variants generated through different helix perturbing modalities do not show as strong a trend, suggesting that while helicity is important it is not the only property that makes a protein protective during desiccation. These results provide direct evidence for the decades old theory that helicity of desiccation‐related IDPs is linked to their anhydrobiotic capacity.This article is protected by copyright. All rights reserved.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 3 more Smart Citations

Helicity of a tardigrade disordered protein contributes to its protective function during desiccation

Biswas,

Gollub,

et al. 2024

Protein Science

View full text Add to dashboard Cite

show abstract

Helicity of a tardigrade disordered protein promotes desiccation tolerance

Biswas

Gollub

et al. 2023

Preprint

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In order to survive extreme drying (anhydrobiosis), many organisms, spanning every kingdom of life, accumulate intrinsically disordered proteins (IDPs). For decades, the ability of anhydrobiosis-related IDPs to form transient amphipathic helices has been suggested to be important for promoting desiccation tolerance. However, evidence empirically supporting the necessity and/or sufficiency of helicity in mediating anhydrobiosis is lacking. Here we demonstrate that the linker region of CAHS D, a desiccation-related IDP from tardigrades that contains significant helical structure, is the protective portion of this protein. Perturbing the sequence composition and grammar of the linker region of CAHS D, through the insertion of helix-breaking prolines, modulating the identity of charged residues, sequence scrambling, or replacement of hydrophobic amino acids with serine or glycine residues results in variants with different degrees of helical structure. Importantly, the resulting helicity of these variants generated through similar helix breaking modalities correlates strongly with their ability to promote desiccation tolerance, providing direct evidence that helical structure is necessary for robust protection conferred by this desiccation-related IDP. However, correlation of protective capacity and helical content in variants generated through different helix perturbing modalities do not show as strong a trend, suggesting that while helicity is important it is not the only property that makes a protein protective during desiccation. These results provide direct evidence for the decades old theory that helicity of desiccation-related IDPs is linked to their anhydrobiotic capacity.

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Structural adaptability and surface activity of tardigrade-inspired peptides

Giubertoni,

Chagri,

Argudo

et al. 2023

Preprint

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Tardigrades are unique micro-animals that withstand harsh conditions, such as extreme temperatures and desiccation. Recently, it was found that specific cytoprotective proteins are essential for ensuring this high environmental tolerance. In particular, cytoplasmic abundant heat soluble (CAHS) proteins, which are intrinsically disordered, adopt more ordered conformations upon desiccation, and are involved in the vitrification of the cytoplasm. The design and synthesis short peptides capable of mimicking the structural behavior (and thus the cytoprotective properties) of CAHS proteins would be beneficial for potential biomedical applications, including the development of novel heat-resistant preservatives for sensitive drug formulations. As a first step in this direction, we selected several model peptides of varying lengths derived from the conserved CAHS motifs 1 and 2, which are part of the intrinsically disordered CAHS-c2 region. We then studied their structures using circular dichroism and linear and two-dimensional infrared spectroscopy in the presence of the desolvating agent TFE (2,2,2-trifluoroethanol), which mimics desiccation. We found that the CAHS model peptides are mostly disordered at 0% TFE (a result that we confirmed by molecular dynamics simulations), but adopt a more 𝛼-helical structure upon the addition of the desolvating agent, similar to what is observed for full CAHS proteins. Additionally, we employed sum frequency generation to investigate the surface activity of the peptides at the air/water interface to mimic a partial dehydration effect. Interestingly, all model peptides are surface active and also adopt a helical structure at the air/water interface. Thus, the selected sequences represent promising model peptides that show similarities in the physicochemical behavior to full CAHS proteins. Our results also suggest that arginine might be a crucial element in defining the strong propensity of these peptides to adopt a helical structure. In the future, the use CAHS model peptides to design new synthetic peptide-based materials could make it possible to mimic and exploit the cytoprotective properties of naturally occurring tardigrade proteins.

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Labile assembly of a tardigrade protein induces biostasis

Cited by 4 publications

References 104 publications

Helicity of a tardigrade disordered protein contributes to its protective function during desiccation

Helicity of a tardigrade disordered protein contributes to its protective function during desiccation

Helicity of a tardigrade disordered protein promotes desiccation tolerance

Structural adaptability and surface activity of tardigrade-inspired peptides

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