.png){kind=icon}
Return
Engineering Functional Membrane-Membrane Interfaces by InterSpy
.png){kind=icon}
OA
AI
DOI：10.1002/smll.202202104.png)
.png){kind=icon}
.png){kind=icon}
Share.png){kind=icon}
Save
Abstract
En 中文
Engineering synthetic interfaces between membranes has potential applications in designing non-native cellular communication pathways and creating synthetic tissues. Here, InterSpy is introduced as a synthetic biology tool consisting of a heterodimeric protein engineered to form and maintain membrane-membrane interfaces between apposing synthetic as well as cell membranes through the SpyTag/SpyCatcher interaction. The inclusion of split fluorescent protein fragments in InterSpy allows tracking of the formation of a membrane-membrane interface and reconstitution of functional fluorescent protein in the space between apposing membranes. First, InterSpy is demonstrated by testing split protein designs using a mammalian cell-free expression (CFE) system. By utilizing co-translational helix insertion, cell-free synthesized InterSpy fragments are incorporated into the membrane of liposomes and supported lipid bilayers with the desired topology. Functional reconstitution of split fluorescent protein between the membranes is strictly dependent on SpyTag/SpyCatcher. Finally, InterSpy is demonstrated in mammalian cells by detecting fluorescence reconstitution of split protein at the membrane-membrane interface between two cells each expressing a component of InterSpy. InterSpy demonstrates the power of CFE systems in the functional reconstitution of synthetic membrane interfaces via proximity-inducing proteins. This technology may also prove useful where cell-cell contacts and communication are recreated in a controlled manner using minimal components.
Keywords:
cell-free expression
membrane protein reconstitution
membrane-membrane interfaces
split-protein reconstitution
synthetic biology
AI Summary
Key information extracted from the uploaded paper, including a brief overview, abstract, background, key highlights, visual analysis, and future outlook.
Journal
IF:
12.1
Papers:
3.0W
Citations:
16.4W
Organization
Cited Papers
Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates.png)
NATURE GENETICS
NATURE GENETICSIF31.8
PREAI
ShareSaveShareSaveEngineering biosynthetic excitable tissues from unexcitable cells for electrophysiological and cell therapy studiesNATURE COMMUNICATIONSOAAI
NATURE COMMUNICATIONSIF15.7
OAAIShareSaveA split fluorescent reporter with rapid and reversible complementationNATURE COMMUNICATIONSOAAI
NATURE COMMUNICATIONSIF15.7
OAAIShareSaveShareSave
ShareSaveShareSave
View more