De novo engineering of intracellular condensates using artificial disordered proteins

Resilin-mimetics as a smart biomaterial platform for biomedical applications

“…90,91 , can potentially lead us a step closer to clinical and therapeutic realization. Moreover, systematic study of programming molecular self-assembly in modular RLP-based systems by extending the design rules applied for IDPs, such as combining two peptide properties into one sequence by increasing fraction of doping and varying block length, and/or blending at varying volume fractions can provide a new platform for design and application of RLPbased materials for intracellular material manipulation 34,92 . In addition, introduction of non-canonical amino acids, such as 4Sfluoroproline and thiazolidine carboxylic acid (isostructural analogues of proline) 93 , and/or post-translational modifications, such as conversion of proline to hydroxyproline, and tyrosine to DOPA 94 could also be potentially applied to expand this design space (increase in functional diversity and development of new RLP-based hybrid biomaterials) with useful properties for applications in nanobiotechnology and medicine.…”

“…The capability of some proteins and polypeptides to change their conformations and self-assemble in response to one or more external stimuli, such as pH, temperature, ions, etc., is encoded in the amino acid sequence level through the types of interactions they are involved 33 , and at the macromolecule level through chain length 34 . The naturally occurring amino acids can undergo various types of interactions including covalent, electrostatic, steric, hydrophobic, π-π stacking and hydrogen bonding.…”

“…Furthermore, with negative zeta potential measured at neutral pH and lack of phase behaviour at pH 12 for Rec1-resilin, it is evident that additional residue interactions, such as cation-π interactions (between arginine (R) and aromatic residues) significantly influence UCST 32,36 . The UCST has been observed to increase with cation-π interactions in the order of tryptophan (W) >> tyrosine (Y) > phenylalanine (F) >> histidine (H) 34 . In addition, the dynamics of phase separation of polypeptides were also observed to be controlled by design parameters, such as increase in the ratio of aromatic:aliphatic residues, RLP repeats or length or molecular weight, and average hydropathy index (AHI) increases UCST 32,34 .…”

“…The UCST has been observed to increase with cation-π interactions in the order of tryptophan (W) >> tyrosine (Y) > phenylalanine (F) >> histidine (H) 34 . In addition, the dynamics of phase separation of polypeptides were also observed to be controlled by design parameters, such as increase in the ratio of aromatic:aliphatic residues, RLP repeats or length or molecular weight, and average hydropathy index (AHI) increases UCST 32,34 . Conversely, the UCST behaviour exhibited by An16, which does not contain the zwitterionic pattern like Rec1-resilin and lysine/ arginine (for cation-π interaction), with no change in zeta potential is quite intriguing and needs further systematic investigation 29 .…”

Cascade communication in disordered networks of enzyme-loaded microdroplets

“…enzymes, within cells or subcellular organelles. 3,4 Spatial separation and positional assembly is to a large extend responsible for the regulation and associated modulation of chemical reactivity, for minimizing the diffusion pathways of intermediates between enzymes, and for the overall efficiency and specificity of biological reactions. [5][6][7][8][9] Also, rivalling reaction centers have to be spatially separated from one another in order to control their destructive interference.…”