Intrinsically disordered proteins (IDP) are proteins whose native state remains unfolded. These proteins are ubiquitous in plants where they play important roles in development and abiotic stress resistance. IDPs stabilize membranes and proteins during periods of water stress such as drought and freezing. The xeroprotective and cryoprotective properties of plant IDPs make them attractive cargoes for delivery into cells as potential solvent-free cellular preservatives. Furthermore, IDP functionality makes them highly amenable to nanoparticle conjugation strategies that would normally abrogate ordered protein bioactivity. For these reasons, we non-covanlently conjugated the cryoprotective <i>Arabidopsis </i>IDPs COR15B and LEA11 to single walled carbon nanotubes (SWNT) to generate aqueous colloidal suspensions of IDP bound to SWNT (IDP-SWNT). We show that the IDPs remain cryoprotective while conjugated to SWNT, are protected from enzymatic digestion by proteases when conjugated to SWNT, and are able to be delivered into HEK293T cells to a higher degree than the free protein. These results lay the groundwork for developing IDP-SWNT as novel nanoparticle-based cellular cryoprotectants. The generalizability of non-covalent SWNT attachment combined with the great diversity of IDPs as functional nanoparticle ligands generates numerous possibilities for use of IDP-SWNT conjugates as cryoprotectants for biological systems.

Intracellular delivery of cryoprotective intrinsically disordered proteins with single-walled carbon nanotubes

ACS Spring Meeting 2021

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