by Tokyo Institute of Technology Bundled filaments were produced from the crystals via an oxidative stress response of cysteinyl thiols after isolation of the crystals from living cells
This method will be considered a useful advance in nano-bio material science and supramolecular chemistry as a synthesis method for constructing sustainable assemblies using protein crystals
Credit: Tokyo Tech Proteins are undoubtedly some of the most fascinating biomolecules, and they perform many of the functions that (in our eyes) separate life from inanimate matter
Multi-molecular protein assemblies even have large-scale structural functions, as evidenced by feathers, hair, and scales in animals
It should come as no surprise that, with progress in advanced nanotechnology and bioengineering, artificial protein assemblies have found applications in a variety of fields, including catalysis, molecular storage, and drug delivery systems
However, producing ordered protein assemblies remains challenging
It is particularly difficult to get monomers, the building blocks of proteins, to assemble stably into the desired structures; this generally requires very accurate design and control of synthesis conditions, such as pH (acidity) and temperature
Recent studies found ways to circumvent this problem by using protein crystals—solid molecular arrangements that occur naturally in some organisms—as precursor matrices to produce protein assemblies
At Tokyo Institute of Technology, Japan, a team of scientists led by Professor Takafumi Ueno has been working on a promising approach for synthesizing protein assemblies from protein crystals
Their strategy involves introducing mutations into the genetic code of an organism that naturally produces protein crystals
These mutations cause disulfide bonds (S-S) to form between monomers in very specific locations in the crystals
The crystals are then dissolved, but instead of breaking down completely into their individual monomers as usual, the newly introduced S-S bonds hold groups of monomers together and the crystals split into many of the desired protein assemblies
With this approach, Ueno's team has managed to synthesize protein cages and tubes by essentially using living cells as nano-3D printers
In their latest study, which was published in Angewandte Chemie International Edition, the team demonstrated yet another application of their novel strategy; this time for the synthesis of bundled protein filaments
They used a culture of insect cells (Spodoptera frugiperda) infected with a virus that caused overexpression of a monomer called "TbCatB
" These monomers naturally aggregate inside the cells into protein crystals, which are held together there by the relatively weak non-covalent interactions between monomers
The scientists strategically introduced two mutations in the cells so that each monomer had two thiol groups (-SH) of cysteine at critical interface points with other monomers
The crystals were extracted from the cells and left to oxidize at room temperature, which caused the thiol groups to change into strong S-S bonds between monomers adjacent along a single direction by autoxidation under air
When the crystals were dissolved, these disulfide bonds, together with some lingering non-covalent interactions, resulted in the formation of bundled protein filaments that were two monomers wide—about 8
3 nanometers
"With our strategy, we achieved a highly precise arrangement of protein molecules while suppressing random aggregation of monomers due to unwanted sulfide bonds, all in a relatively straightforward one-pot process," highlights Ueno
Overall, the approach demonstrated by the team at Tokyo Tech stands as an innovative way to synthesize protein structures via rational genetic engineering and by using the tools naturally available to cells of certain organisms
"We consider our synthesis method a useful advance in nano-biomaterials science and supramolecular chemistry for producing desired stable assemblies from protein crystals," concludes Ueno
Only time will tell what other useful molecular structures can be produced using this strategy and what interesting applications they will find
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