Synthesis and Evaluation of Cereblon‐Recruiting HaloPROTACs

Synthesis and Evaluation of Cereblon-Recruiting HaloPROTACs

A bifunctional HaloPROTAC ligand was developed to recruit the CRBN E3 ubiquitin ligase to degrade target proteins fused with a HaloTag in the cell. The CRBN-recruiting HaloPROTAC can expand the use of the HaloPROTAC system for screening the degradation targets for therapeutic discovery.


Abstract

Target validation is key to the development of protein degrading molecules such as proteolysis-targeting chimeras (PROTACs) to identify cellular proteins amenable for induced degradation by the ubiquitin-proteasome system (UPS). Previously the HaloPROTAC system was developed to screen targets of PROTACs by linking the chlorohexyl group with the ligands of E3 ubiquitin ligases VHL and cIAP1 to recruit target proteins fused to the HaloTag for E3-catalyzed ubiquitination. Reported here are HaloPROTACs that engage the cereblon (CRBN) E3 to ubiquitinate and degrade HaloTagged proteins. A focused library of CRBN-pairing HaloPROTACs was synthesized and screened to identify efficient degraders of EGFP-HaloTag fusion with higher activities than VHL-engaging HaloPROTACs at sub-micromolar concentrations of the compound. The CRBN-engaging HaloPROTACs broadens the scope of the E3 ubiquitin ligases that can be utilized to screen suitable targets for induced protein degradation in the cell.

Effects of Replacing a Central Glycine Residue in GLP‐1 on Receptor Affinity and Signaling Profile

Effects of Replacing a Central Glycine Residue in GLP-1 on Receptor Affinity and Signaling Profile

The receptor activated by GLP-1 is a target for drugs that treat type 2 diabetes and obesity. Since glycine has a low α-helix propensity, and Gly22 of GLP-1 does not contact the receptor, we replaced Gly22 with residues that have a higher or lower helix propensity. Most substitutions had little effect on receptor affinity or activation and raised the possibility that the receptor-bound state of agonist peptides encompasses a broader conformational envelope.


Abstract

Agonists of the glucagon-like peptide-1 receptor (GLP-1R) are used to treat diabetes and obesity. Cryo-EM structures indicate that GLP-1 is completely α-helical when bound to the GLP-1R. The mature form of this hormone, GLP-1(7-36), contains a glycine residue near the center (Gly22). Since glycine has the second-lowest α-helix propensity among the proteinogenic α-amino acid residues, and Gly22 does not appear to make direct contact with the receptor, we were motivated to explore the impact on agonist activity of altering the α-helix propensity at this position. We examined GLP-1 analogues in which Gly22 was replaced with L-Ala, D-Ala, or β-amino acid residues with varying helix propensities. The results suggest that the receptor is reasonably tolerant of variations in helix propensity, and that the functional receptor-agonist complex may comprise a conformational spectrum rather than a single fixed structure.

Molecular Highway Patrol for Ribosome Collisions

Molecular Highway Patrol for Ribosome Collisions

Car crashes on highways require responses like rescuing victims, deviating traffic, towing away damaged vehicles, and sometimes even blocking a road. Similarly, during translation, ribosomes can collide into each other with potentially dangerous outcomes for the cell. In response to ribosome collisions, cells mount a multi-pronged response, activating several molecular ′squads′, symbolized by the Highway Police badge. These include degradation pathways for faulty mRNAs, peptide products and sometimes ribosomal constituents, regulation of translation, and stress as well as immune responses. More information can be found in the Review by M.-L. Winz et al.


Synthesis of Short Peptides with Perfluoroalkyl Side Chains and Evaluation of Their Cellular Uptake Efficiency

Synthesis of Short Peptides with Perfluoroalkyl Side Chains and Evaluation of Their Cellular Uptake Efficiency

We have synthesized perfluoroalkyl-containing amino acids (RF-AA) and demonstrated that tripeptides composed of the RF-AA have high cellular uptake efficiency. Furthermore, the absolute configuration of RF-AA affected the uptake efficiency due to the change in its ability to form nanoparticles. We anticipate that these findings provide the foundation for the development of efficient cell-penetrating peptide (CPP). The cover picture shows tripeptides with perfluoroalkyl groups labeled with hydrophilic dyes that form particles in water and are efficiently taken up into cells. More information can be found in the Research Article by K. Aikawa et al.


Self‐consistent field method for open‐shell systems within the density‐matrix functional theory

Abstract

The unrestricted Hartree-Fock method is extended to correlation calculation within the density-matrix functional theory. The method is derived from an entropic cumulant functional for the correlation energy. The eigenvalue equations for the spin-orbitals are modified by the orbital occupation numbers. The Euler equation for the occupation numbers results in the Fermi-Dirac distribution, which is very efficient to update as soon as the orbital eigenvalue equations are solved. The method is demonstrated on the ground state of O2$$ {}_2 $$.

Emergence of ATP‐ and GTP‐Binding Aptamers from Single RNA Sequences by Error‐Prone Replication and Selection

Emergence of ATP- and GTP-Binding Aptamers from Single RNA Sequences by Error-Prone Replication and Selection**

Function in RNA oligomers - specifically low μM specific binding of purine (ATP and GTP) nucleotides - can arise from single, non-functional and partially homopolymeric (polyA) sequences of minimal structural and informational complexity by iterative cycles of mutation and selection.


Abstract

The spontaneous emergence of function from diverse RNA sequence pools is widely considered an important transition in the origin of life. Here we show that diverse sequence pools are not a prerequisite for the emergence of function. Starting five independent selection experiments each from a single RNA seed sequence - comprising a central homopolymeric poly-A (or poly-U) segment flanked by different conserved primer binding sites - we observe transformation (continuous drift) of the seeds into low diversity sequence pools by mutation, truncation and recombination without ever reaching that of a random pool even after 24 rounds. Upon continuous error prone replication and selection for ATP binding we isolate specific ATP- or GTP-binding aptamers with low micromolar affinities. Our results have implications for early RNA evolution in the light of the high mutation rates associated with both non-enzymatic and enzymatic prebiotic RNA replication.

Elucidating the Mechanism of Freeze‐Thaw Driven Content Mixing between Protocells

Elucidating the Mechanism of Freeze-Thaw Driven Content Mixing between Protocells

Freeze-thaw cycles serve as physicochemical driving force for the lateral exchange of enclosed material between giant lipid vesicles. It was demonstrated that this exchange relies on transient periods of membrane permeability leading to content diffusion across vesicle membranes. Moreover, we explored and quantified essential parameters affecting the lateral transfer efficiency.


Abstract

Modern cells rely on highly evolved protein networks to accomplish essential life functions, including the inheritance of information from parents to their offspring. In the absence of these sophisticated molecular machineries, alternatives were required for primitive protocells to proliferate and disseminate genetic material. Recurring environmental constraints on ancient earth, such as temperature cycles, are considered as prebiotically plausible driving forces capable of shuffling of protocellular contents, thereby boosting compositional complexity. Using confocal fluorescence microscopy, we show that temperature oscillations such as freezing-thawing (FT) cycles promote efficient content mixing between giant unilamellar vesicles (GUVs) as model protocells. We shed light on the underlying exchange mechanism and demonstrate that transient periods of destabilized membranes enable the diffusion of cargo molecules across vesicle membranes. Furthermore, we determine essential parameters, such as membrane composition, and quantify their impact on the lateral transfer efficiency. Our work outlines a simple scenario revolving around inter-protocellular communication environmentally driven by periodic freezing and melting of water.

Influence of Silicate Rock Glass Compositions on the Efficacy of Prebiotic RNA Polymerization Reactions: The Case of 3’,5’ Cyclic Guanosine Monophosphate

Influence of Silicate Rock Glass Compositions on the Efficacy of Prebiotic RNA Polymerization Reactions: The Case of 3’,5’ Cyclic Guanosine Monophosphate

Glass composition matters: This work demonstrates that the outcome of dry-phase prebiotic polymerization reactions could have been strongly affected by the oxide composition of the hosting rock glasses.


Abstract

Silicate glasses are ubiquitous on terrestrial planets wherever molten rock – generated by volcanism or impacts – is quenched by air or water. Hence, they provided ready and abundant reactive solid surfaces for prebiotic chemistry on the early Earth. Here, we show that rock glass composition determines basicity, which in turn modulates the outcome of prebiotic synthetic processes such as the polymerization of 3’, 5’ cyclic guanosine monophosphate.

Front Cover: Elucidating the Mechanism of Freeze‐Thaw Driven Content Mixing between Protocells (ChemSystemsChem 5/2023)

Front Cover: Elucidating the Mechanism of Freeze-Thaw Driven Content Mixing between Protocells (ChemSystemsChem 5/2023)

The Front Cover illustrates the exchange of genetic material between model protocells through cycles of freezing and thawing. Freeze-thaw cycles as prebiotic environmental driver induce a transient increase in membrane permeability enabling the lateral transfer of genetic information in a population of primitive protocells. More information can be found in the Research Article by Benedikt Peter and Petra Schwille.


Molecular Engineering of Carbohydrate Recognition

Molecular Engineering of Carbohydrate Recognition

The recognition of carbohydrates has been of growing interest in the supramolecular chemistry field, with specific relevance for numerous potential applications. At the same time, the binding many of carbohydrates in water selectively and with high affinity still presents a number of practical challenges that inspire ongoing research efforts toward new classes of bio-inspired synthetic receptors.


Abstract

Carbohydrates play a number of structural, functional, and metabolic roles in underpinning natural life processes, acting in states of both health and disease. Given this importance, over millions of years of evolution, living systems have developed an ability to recognize and bind carbohydrates, achieving remarkable recognition affinity and specificity in spite of the often hydrophilic and ubiquitous character of carbohydrate targets. In recent years, bio-inspired synthetic receptors have been developed to bind carbohydrates, with examples of (pseudo)temple-shaped receptors, flexible receptors, and dynamic-covalent/coordinative receptors reported. Certain of these have even demonstrated promising results, for example in binding glucose or exhibiting antiviral and antibiotic function. Accordingly, and in spite of remaining challenges, the development of synthetic receptors for carbohydrate recognition holds great promise to combat some of the most urgent problems facing our world today.