RT Journal Article
SR Electronic
T1 Encoding extracellular modification of artificial cell membranes using engineered self-translocating proteins
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2023.10.06.561148
DO 10.1101/2023.10.06.561148
A1 Harjung, Alexander
A1 Fracassi, Alessandro
A1 Devaraj, Neal
YR 2023
UL http://biorxiv.org/content/early/2023/10/06/2023.10.06.561148.abstract
AB A common method of generating artificial cells is to encapsulate protein expression systems within lipid vesicles. However, to communicate with the external environment, protein translocation across lipid membranes must take place. In living cells, protein transport across membranes is achieved with the aid of complex translocase systems which are difficult to reconstitute into artificial cells. Thus, there is need for simple mechanisms by which proteins can be encoded and expressed inside synthetic compartments yet still be externally displayed. Here we present a genetically encodable membrane functionalization system based on mutants of pore-forming proteins. We show that the membrane translocating loop of α-hemolysin can be engineered to translocate functional peptides up to 52 amino acids across lipid membranes. Engineered hemolysins can be used for genetically programming artificial cells to display interacting peptide pairs, enabling their assembly into artificial tissue-like structures capable of signal transduction.Competing Interest StatementThe authors have declared no competing interest.