Analysis of Biomolecular Condensates in Anhydrobiosis

A few species, such as vermiform rotifers, can survive severe dehydration in a near pseudo-dead state, a phenomenon known as "anhydrobiosis". Anhydrobiosis has evolved many molecular tools and adaptations to achieve this phenomenon. In addition to the accumulation of specific compatible osmolytes and the metabolic down-regulation often observed before water loss, anhydrobiosis utilizes a set of intrinsically disordered proteins (IDPs) to protect cells during osmotic stress and complete desiccation. Late embryonic developmental abundance (LEA) proteins are one of the most important families of polypeptides associated with dehydration. LEA proteins are classified as IDPs and are closely associated with increased desiccation tolerance in plants and many anhydrobiotic animals. It has been shown that some LEA proteins form protein droplets during water loss and these droplets selectively separate biomolecules and may organize cytoplasmic lysates into biochemically distinct refugee regions for dehydration-sensitive biomolecules.

Fig. 1. AfrLEA6 is predicted to have three distinct regions. (Belott C, et al., 2020)

Customized Services

LEA proteins form a large class of intrinsically disordered proteins associated with dehydrated organisms and are widely found in plants, rotifers, nematodes, and arthropods. CD BioSciences offers professional anhydrobiosis biomolecular condensates analysis services to help clients gain a thorough understanding of the molecular principles that control anhydrobiosis and the role of liquid-liquid phase separation (LLPS) of proteins in the embryonic process.

• Bioinformatics Analysis

We offer a variety of bioinformatics tools to classify LEA proteins and assess the structural propensity of Artemia franciscana LEA6 (Afr LEA6), including:

• Identification of amino acid homology regions and internal repeat sequences in the Afr LEA6 protein sequence.
• Modeling the potential secondary and tertiary structural motifs of Afr LEA6.
• Comparison with homologous structures in proteins with known crystal structures.

• Characterization of Afr LEA6 LLPS

We provide multiple technical platforms to analyze the behavior of Afr LEA6 when ectopically expressed in Drosophila under dry stress in Kc167 cells and purified proteins in solution.

• We can construct Afr LEA6 condensate partitioning GFP to analyze the protein surface charge on intermolecular interactions between Afr LEA6 and other proteins.
• We provide confocal microscopy to characterize the behavior of Afr LEA6 during the transition from a low water environment to a fully dry state, as well as to image Kc167 cells expressing both Afr LEA6-mCherry and enhanced GFP (eGFP).

• Anhydrobiosis Biomolecular Condensate Engineering

Our experts are committed to engineering this property of anhydrobiosis into organisms susceptible to water stress. We aim to implant anhydrous bioengineering into other organisms to facilitate transformative advances in everything from cell preservation techniques to improving drought tolerance in crops.

CD BioSciences aims to help clients characterize biomolecular condensates in anhydrobiosis and to analyze the molecular mechanisms by which LLPS promotes desiccation tolerance in animals. If you have any special requirements for our services, please feel free to contact us. We are looking forward to working together on your attractive projects.