LEA_4 motifs function alone and in conjunction with synergistic cosolutes to protect a labile enzyme during desiccation

Protein Science, Год журнала: 2025, Номер 34(2)

Опубликована: Янв. 22, 2025

Abstract Organisms from all kingdoms of life depend on Late Embryogenesis Abundant (LEA) proteins to survive desiccation. LEA are divided into broad families distinguished by the presence family‐specific motif sequences. The LEA_4 family, characterized 11‐residue motifs, plays a crucial role in desiccation tolerance numerous species. However, these motifs function is unclear, with some studies finding that they recapitulate full‐length vivo, and other opposite result. In this study, we characterize ability protect desiccation‐sensitive enzyme, citrate synthase (CS), loss during We show here not only prevent CS but also can do so more robustly via synergistically interactions cosolutes. Our analysis further suggests cosolutes induce synergy manner correlates transfer free energy. This research advances our understanding demonstrating their specific clients varying degrees protective capacity modulated chemical environment. findings extend beyond realm tolerance, offering insights interplay between IDPs By investigating highlight broader strategies for protein stability function.

Язык: Английский

Diversity in the protective role(s) of the conserved motif 1 from tardigrade cytoplasmic‐abundant heat‐soluble proteins during drying

Protein Science, Год журнала: 2025, Номер 34(3)

Опубликована: Фев. 19, 2025

In their recent manuscript, Protective roles of highly conserved motif 1 in tardigrade cytosolic-abundant heat soluble protein extreme environments, Kang et al. (2024) investigate the putative role a cytoplasmic-abundant heat-soluble (CAHS) protein, PrCAHS 1, desiccation protection. To do so, test ability different regions to confer protection desiccation-sensitive enzyme lactate dehydrogenase (LDH). doing this, they compare how mixtures with mass ratios LDH and retain activity after drying rehydration. However, another way comparing protective capacity protectants molecular weights is at similar molar ratios. Since 19-mer has weight that approximately 11 times lower than full-length choices about comparisons (weight vs. comparisons) are made could influence outcome comparison. This led us reanalyze data for converting reported ratios, performing statistical analysis across (Figure 1a,b). reanalysis confirmed finding by outperforms protecting during drying. this result was surprising us, since other proteins from CAHS family provide robust (Biswas al., 2024; Boothby 2017; Hesgrove 2021; Kc Nguyen 2022; Packebush 2023; Piszkiewicz 2019; Sanchez-Martinez 2024) so more efficiently individual domains 2023). observation raises question whether feature among proteins. assess if 1s proteins, we performed set assay those (2024), using an ortholog HeCAHS 8, derived 8. Here use 8 behave similarly 1. We observed opposite obtained where provided LDH, whereas did not 1c,d). From these results, one can infer while some may help enzymatic greater it from, sequences not. suggests functional diversity rather all These results line study found LEA_4 robustly protect drying, motifs (Kc Nicholson 2025). Why might have were al (2024)? One possibility distinct general, truly function differently. possibility, fact sequence 1e), minor differences composition account differential between two motifs. Previous work demonstrated small changes CAHS-inspired peptides lead large structural ensemble Giubertoni 2024), which turn affect mediators 2024). Another possible source discrepancy methods measuring activity. While indirect tetrazolium/formazan assay, opted method directly assays presence NADH (a primary product activity), widely used tolerance field Goyal 2005; Although methodology as been utilized former on LEA protein's (Hatanaka 2013), does necessarily make best approach assay. Several issues exist assays, including propensity formazan precipitate high concentration, thus causing major inconsistencies quantification (Riss 2016). The effect such precipitation concentrations sample levels appear actually low Companies sell often mention include excipients reduce precipitation; however, generally difficult obtain precise information what effectiveness preventing precipitation. explanation why be suggested or domains, prevent aggregation LDH. anti-aggregation potential measure absorbance 340 nm solutions prepared desiccating then rehydrating alone authors first + 1f, replotted They conclude latter solution alone, increased due but itself, contribution readings impossible based data. It worth mentioning here known undergo phase transition form gels concentration-dependent manner, (Eicher Malki Tanaka Yagi-Utsumi 2021). suggest increase gelation conclusion cause well desiccation-related IDPs, thought preserve through prevention aggregation, mechanisms (Chakrabortee 2007; Koubaa 2019). conducted experiments utilizing measured UV-light (340 nm). same making our comparable theirs. size exclusion chromatography (SEC) considered gold standard studying (Hong 2012). because 2021), SEC feasible will clog column absence strong denaturant, also perturb began mixed both before drying/rehydration. Dried/rehydrated samples showed minimal significant rehydration relative hydrated samples. show any upon drying-rehydration. implies itself. Moreover, combination 1g). Finally, arithmetic addition value (alone) compared (LDH 8) shows decrease dried state actual mixture 1h). If would expect see statistically sum plus alone. observe unit components. that, helps 1h) 1c). Using composed report consisting just no significant. authors, one-way ANOVA Tukey's post-hoc 1f). revealed there difference Furthermore, BSA indicates PrCAHS1 (or inducing) interpretation emphasizes importance tests support conclusions drawn experiment, unfortunately lacking (2024). conclusion, portions specifically helical linker region, previously shown enzymes 2023), evidence process lacking. isolation Our data, alongside mitigating drying-induced dysfunction. further like caution when analyzing IDP functionality, important consider extending relevance observations partial native context problematic. A fragment take conformational differ its conformation Das 2015). exacerbated under environmental conditions impart IDPs acknowledge done hope commentary contribute fruitful discussion biology tardigrades fascinating Sourav Biswas: Conceptualization; investigation; writing – original draft; methodology; visualization; review editing; formal analysis; curation. Thomas C. Boothby: editing. thank Dr. Chin-Ju Park (Gwangju Institute Science Technology) providing raw associated manuscript here. declare conflict interest. findings available corresponding author reasonable request.

Язык: Английский

Protein surface chemistry encodes an adaptive tolerance to desiccation

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июль 29, 2024

Abstract Cellular desiccation - the loss of nearly all water from cell is a recurring stress in an increasing number ecosystems that can drive protein unfolding and aggregation. For cells to survive, at least some proteome must resume function upon rehydration. Which proteins tolerate desiccation, molecular determinants underlie this tolerance, are largely unknown. Here, we apply quantitative structural proteomic mass spectrometry show certain possess innate capacity rehydration following extreme loss. Structural analysis points surface chemistry as key determinant for which test by showing rational mutants convert sensitive into tolerant one. Desiccation tolerance also has strong overlap with cellular function, highly responsible production small molecule building blocks, intolerant involved energy-consuming processes such ribosome biogenesis. As result, rehydrated preferentially enriched metabolite producers depleted cell’s heaviest consumers. We propose functional bias enables kickstart their metabolism promote survival Teaser Proteins resist dryness tuning amino acids on surfaces.

Язык: Английский