Author Correction: Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry

Abstract

The previously published version of this Article contained a consequential error in the fourth sentence of the "Discussion", which incorrectly read 'The use of the azF-SCO crosslink proved important as classical disulfide crosslinking did improve protein function'. The correct version states 'The use of the azF-SCO crosslink proved important as classical disulfide crosslinking did not improve protein function'-the word 'not' has been added as the 14th word. This has been corrected in both the PDF and HTML versions of the Article.

Better together: building protein oligomers naturally and by design

“…In the case above, the original monomeric proteins are largely inactive that are switched on when dimerised through linkage via residue 148 (green spheres, PDB 5NHN). Figure is copied with permission from Worthy et al [62] under a creative commons license. ( b ) Scheme showing the oligomerisation of a synthetic ankyrin repeat protein (grey), containing the metal-chelating ncAA 2,2′-bipyridin-5yl-alanine (Bpy-ala, shown as sticks).…”

“…The Jones group has successfully demonstrated the use of genetically encoded SPAAC by linking monomers of fluorescent proteins together to create functionally enhanced dimers of super-folder green fluorescent protein (sfGFP) (Figure 2a) [62]. The fluorescence mechanism and properties of sfGFP and other Aequorea victoria derived FPs is well defined [63–66] with the central active component being the solvent-shielded chromophore, buried within the β-barrel structure.…”

“…Mutation of H148 to a ncAA results in the breakage of this key H-bond causing the CRO A chromophore to predominate [51,54]. The formation of sfGFP homodimers using SPAAC compatible residues at 148 not only reverses this protonation state so switching on CRO B, but enhances brightness over threefold above wild type sfGFP — indicative of functional synergy [62]. Comparison of the 400 nm : 485 nm excitation peaks would thus allow a ratiometric estimation of the CRO A monomer to CRO B dimer population.…”

“…Attaching these FPs to separate interacting proteins brings the complex together to form a FP dimer and so enhancing the fluorescence output and thus the SNR. However, considering the aforementioned developments in mutations centred on residue 148 of sfGFP mutants [51,62], it is exciting to note the potential homodimers may have used as a novel suite of ddFP biosensors. Not only will these be easier to engineer than heterodimers, homodimers are likely to be more stable to fluctuations in the microenvironment.…”

Metal-Mediated Protein Assembly Using a Genetically Incorporated Metal-Chelating Amino Acid

“…Although a protein monomer can function independently, proteins often form complex structures to carry out sophisticated functions. Since protein complexation enables proteins to enhance their performance, researchers have attempted to construct artificial protein complexes and protein–protein conjugates. − DNA nanotechnology is relatively well-established, and DNA assembly systems have been developed to form complex and defined nanostructures using Watson–Crick base-pairing . On the other hand, protein nanotechnology has been recently developed and is a growing research field; this is because proteins consist of twenty building blocks and form defined structures through complex molecular interactions, while DNA consists of four building blocks and uses simple and general rules in their interactions.…”