Cell-free co-synthesis of protein nanoassemblies: tubes, rings, and doughnuts.
We used a cell-free transcription/translation system to synthesize structural proteins of the T4 bacteriophage. We focused on two proteins that participate in the formation of the virus tail tube assembly. Synthesized separately, the proteins assembled into their in vivo forms, namely one polymerized into rigid hollow nanotubes approximately 20 nm thick and hundreds of nanometers long, the other assembled into 10 nm tube-capping hexameric rings. Co-synthesis of the two proteins, however, revealed a novel structure of a nanodoughnut with an outer diameter of approximately 50 nm and thickness of approximately 20 nm. Cell-free co-synthesis and assembly of T4 structural proteins can be extended in a combinatorial fashion. The addition of other structural genes offers control of native nanoassemblies and may reveal ones not observable by mixing purified components.
Daube
Shirley S
SS
Kimmel Center for Nanoscale Science, Chemical Research Support, Department of Materials and Interfaces, The Weizmann Institute of Science, Rehovot, Israel 76100.
Arad
Talmon
T
Bar-Ziv
Roy
R
D017122:Bacteriophage T4 / Q000378:metabolism / Q000648:ultrastructure; D002474:Cell-Free System; D008854:Microscopy, Electron; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D036103:Nanotechnology; D045333:Nanotubes, Peptide / Q000737:chemistry* / Q000648:ultrastructure; D017376:Viral Tail Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000648:ultrastructure
D045333:Nanotubes, Peptide; D017376:Viral Tail Proteins; C402835:glycoprotein 18, bacteriophage T4