Rational Design and Self-Assembly of Coiled-Coil Linked
SasG Protein Fibrils
Posted on 2020-06-22 - 11:37
Protein engineering is an attractive
approach for the self-assembly
of nanometer-scale architectures for a range of potential nanotechnologies.
Using the versatile chemistry provided by protein folding and assembly,
coupled with amino acid side-chain functionality, allows for the construction
of precise molecular “protein origami” hierarchical
patterned structures for a range of nanoapplications such as stand-alone
enzymatic pathways and molecular machines. The Staphyloccocus
aureus surface protein SasG is a rigid, rod-like structure
shown to have high mechanical strength due to “clamp-like”
intradomain features and a stabilizing interface between the G5 and
E domains, making it an excellent building block for molecular self-assembly.
Here we characterize a new two subunit system composed of the SasG
rod protein genetically conjugated with de novo designed
coiled-coils, resulting in the self-assembly of fibrils. Circular
dichroism (CD) and quartz-crystal microbalance with dissipation (QCM-D)
are used to show the specific, alternating binding between the two
subunits. Furthermore, we use atomic force microscopy (AFM) to study
the extent of subunit polymerization in a liquid environment, demonstrating
self-assembly culminating in the formation of linear macromolecular
fibrils.
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Jasaitis, Lukas; Silver, Callum D.; Rawlings, Andrea E.; Peters, Daniel T.; Whelan, Fiona; Regan, Lynne; et al. (2020). Rational Design and Self-Assembly of Coiled-Coil Linked
SasG Protein Fibrils. ACS Publications. Collection. https://doi.org/10.1021/acssynbio.0c00156
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AUTHORS (10)
LJ
Lukas Jasaitis
CS
Callum D. Silver
AR
Andrea E. Rawlings
DP
Daniel T. Peters
FW
Fiona Whelan
LR
Lynne Regan
LP
Laia Pasquina-Lemonche
JP
Jennifer R. Potts
SJ
Steven D. Johnson
SS
Sarah S. Staniland
KEYWORDS
SasG rod proteinrod-like structureE domainsSasG Protein Fibrils Protein engineeringacid side-chain functionalitybuilding blocknanometer-scale architecturesself-assemblyRational Designforce microscopyAFMQCM-DStaphyloccocus aureus surface protein SasGG 5Circular dichroismsubunit polymerizationfibrilquartz-crystal microbalancesubunit system