Fine‐Tuning Protein Self‐Organization by Orthogonal Chemo‐Optogenetic Tools

Sun, Huan; Jia, Haiyang; Ramirez-Diaz, Diego A.; Budiša, Nediljko; Schwille, Petra

doi:10.1002/ange.202008691

Cited by 4 publications

(3 citation statements)

References 34 publications

(36 reference statements)

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“…The position is located on the outer surface of the protein and is exposed to solvent, allowing us to mimic natural halogenation progress that is modified by external natural modifiers. In addition, this position is known to be sensitive to PTMs 26 , 46 , making it a good candidate for investigation of halogenations.…”

Section: Resultsmentioning

confidence: 99%

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Halogenation of tyrosine perturbs large-scale protein self-organization

et al. 2022

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Protein halogenation is a common non-enzymatic post-translational modification contributing to aging, oxidative stress-related diseases and cancer. Here, we report a genetically encodable halogenation of tyrosine residues in a reconstituted prokaryotic filamentous cell-division protein (FtsZ) as a platform to elucidate the implications of halogenation that can be extrapolated to living systems of much higher complexity. We show how single halogenations can fine-tune protein structures and dynamics of FtsZ with subtle perturbations collectively amplified by the process of FtsZ self-organization. Based on experiments and theories, we have gained valuable insights into the mechanism of halogen influence. The bending of FtsZ structures occurs by affecting surface charges and internal domain distances and is reflected in the decline of GTPase activities by reducing GTP binding energy during polymerization. Our results point to a better understanding of the physiological and pathological effects of protein halogenation and may contribute to the development of potential diagnostic tools.

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Section: Resultsmentioning

confidence: 99%

“…1a ). The protein has been shown to be sensitive to halogenating chemicals 25 and its activity is sensitive to modifications at individual sites 26 . As an essential part of the bacterial division ring, known as “Z ring”, FtsZ has shown intriguing self-organization when reconstituted in vitro on biological membranes.…”

Section: Introductionmentioning

confidence: 99%

Halogenation of tyrosine perturbs large-scale protein self-organization

et al. 2022

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“…Furthermore, the complete orthogonality of T7RNAp to all endogenous RNA polymerases in prokaryotic and eukaryotic systems allows for the photochemical activation of gene expression in the cells. Similarly, Sun and colleagues 456 replaced Tyr222 with ONBY (27) into the prokaryotic filamentous cell-division protein (FtsZ), which is highly important for the assembly of the division ring in bacteria. Though masking Tyr222 still shows self-assembly into filaments, dynamic self-organization into ring patterns was abolished.…”

Section: Stop Codon Suppressionmentioning

confidence: 99%

Recent Advances in Biocatalysis with Chemical Modification and Expanded Amino Acid Alphabet

et al. 2021

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The two main strategies for enzyme engineering, directed evolution and rational design, have found widespread applications in improving the intrinsic activities of proteins. Although numerous advances have been achieved using these ground-breaking methods, the limited chemical diversity of the biopolymers, restricted to the 20 canonical amino acids, hampers creation of novel enzymes that Nature has never made thus far. To address this, much research has been devoted to expanding the protein sequence space via chemical modifications and/or incorporation of noncanonical amino acids (ncAAs). This review provides a balanced discussion and critical evaluation of the applications, recent advances, and technical breakthroughs in biocatalysis for three approaches: (i) chemical modification of cAAs, (ii) incorporation of ncAAs, and (iii) chemical modification of incorporated ncAAs. Furthermore, the applications of these approaches and the result on the functional properties and mechanistic study of the enzymes are extensively reviewed. We also discuss the design of artificial enzymes and directed evolution strategies for enzymes with ncAAs incorporated. Finally, we discuss the current challenges and future perspectives for biocatalysis using the expanded amino acid alphabet.

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Photoactivation of Innate Immunity Receptor TLR8 in Live Mammalian Cells by Genetic Encoding of Photocaged Tyrosine

et al. 2021

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The effectiveness of innate immune responses relies on an intricate balance between activation and regulation. TLR8, a member of the Toll‐like receptor (TLR) family, plays a fundamental role in host defense by sensing viral single‐stranded RNAs (ssRNAs). However, the molecular recognition and regulatory mechanism of TLR8 is not fully understood, especially in a whole‐cell environment. Here, we engineer the first light‐controllable TLR8 model by genetically encoding a photocaged tyrosine, NBY, into specific sites of TLR8. In the caged forms, the activity of TLR8 is masked but can be restored upon decaging by exposure to UV light. To explain the mechanism clearly, we divide the sites with light responsiveness into three groups. They can separately block the ligands that bind to the pockets of TLR8, change the interaction modes between two TLR8 protomers, and interfere with the interactions between TLR8 cytosolic domains with its downstream adaptor. Specifically, we use this chemical caging strategy to probe and evaluate the function of several tyrosine sites located at the interface of TLR8 homodimers with a previously unknown regulatory mode, which may provide a new strategy for TLR8 modulator development. Effects on downstream signaling pathways are monitored at the transcriptional and translational levels in various cell lines. By photoactivating specific cells within a larger population, this powerful tool can provide novel mechanistic insights, with potential in biotechnological and pharmaceutical applications.

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Fine‐Tuning Protein Self‐Organization by Orthogonal Chemo‐Optogenetic Tools

Cited by 4 publications

References 34 publications

Halogenation of tyrosine perturbs large-scale protein self-organization

Halogenation of tyrosine perturbs large-scale protein self-organization

Recent Advances in Biocatalysis with Chemical Modification and Expanded Amino Acid Alphabet

Photoactivation of Innate Immunity Receptor TLR8 in Live Mammalian Cells by Genetic Encoding of Photocaged Tyrosine

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