Biophysical Insights into Drug Discovery: Leveraging Phase Transitions and Protein Behavior for Therapeutic Innovation DOI
Afam Uzorka,

David Kibirige,

David Makumbi

et al.

Biophysical Reviews and Letters, Journal Year: 2023, Volume and Issue: 18(03), P. 171 - 176

Published: Sept. 1, 2023

In the search for novel treatments various diseases, nexus of biophysics and drug discovery represents a dynamic transformational paradigm. A new age in pharmaceutical research has begun thanks to its thorough grasp structural physical characteristics biological molecules. This study explores how advances biophysical approaches, including allosteric modulation, intrinsically disordered proteins (IDPs), protein phase transitions, have transformed development process. Protein supported by principles, provided crucial insights into disorders like ALS Alzheimer’s, opening up avenues therapeutic intervention. Targeting IDPs their role liquid–liquid separation produced creative approaches diseases that were formerly thought be resistant therapy. order reduce complexity complicated concept made possible understanding, offers precise selective approach medication creation. review highlights significant influence on therapeutics drugs. Biophysics is advancing discipline toward creation more precise, efficient, tailored medications illuminating properties proteins, complexities dynamics interactions. Biophysical are poised alter healthcare bringing together several fields sustained innovation, giving people dealing with variety ailments fresh hope.

Language: Английский

Cracking the Code: Reprogramming the Genetic Script in Prokaryotes and Eukaryotes to Harness the Power of Noncanonical Amino Acids DOI

Cosimo Jann,

Sabrina Giofrè,

Rajanya Bhattacharjee

et al.

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(18), P. 10281 - 10362

Published: Aug. 9, 2024

Over 500 natural and synthetic amino acids have been genetically encoded in the last two decades. Incorporating these noncanonical into proteins enables many powerful applications, ranging from basic research to biotechnology, materials science, medicine. However, major challenges remain unleash full potential of genetic code expansion across disciplines. Here, we provide an overview diverse methodologies systems their final applications prokaryotes eukaryotes, represented by

Language: Английский

Citations

16
Evolution of Pyrrolysyl-tRNA Synthetase: From Methanogenesis to Genetic Code Expansion DOI Creative Commons
Nikolaj G. Koch, Nediljko Budiša

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(16), P. 9580 - 9608

Published: July 2, 2024

Over 20 years ago, the pyrrolysine encoding translation system was discovered in specific archaea. Our Review provides an overview of how once obscure pyrrolysyl-tRNA synthetase (PylRS) tRNA pair, originally responsible for accurately translating enzymes crucial methanogenic metabolic pathways, laid foundation burgeoning field genetic code expansion. primary focus is discussion to successfully engineer PylRS recognize new substrates and exhibit higher

Language: Английский

Citations

14
Recent progress of chemical methods for lysine site-selective modification of peptides and proteins DOI
Jian Li, Jinjin Chen, Qi-Long Hu

et al.

Chinese Chemical Letters, Journal Year: 2024, Volume and Issue: unknown, P. 110126 - 110126

Published: June 1, 2024

Language: Английский

Citations

4
One‐pot dual protein labeling for simultaneous mechanical and fluorescent readouts in optical tweezers DOI Creative Commons
Laura‐Marie Silbermann, Maximilian Fottner, Ronald van der Meulen

et al.

Protein Science, Journal Year: 2025, Volume and Issue: 34(4)

Published: March 18, 2025

Abstract Optical tweezers are widely used in the study of biological macromolecules but limited by their one‐directional probing capability, potentially missing critical conformational changes. Combining fluorescence microscopy with optical tweezers, employing Förster resonance energy transfer (FRET) pairs, addresses this issue. When integrating orthogonal protein conjugation methods needed to enable simultaneous, site‐specific attachment fluorophores and DNA handles, commonly apply force molecules interest. In study, we utilized commercially available reagents for dual labeling homodimeric heat shock 90 (Hsp90) using thiol‐maleimide inverse electron demand Diels–Alder cycloaddition (IEDDAC) bioorthogonal reactions. a one‐pot approach, Hsp90 modified cysteine mutation non‐canonical amino acid cyclopropene‐L‐lysine (CpK) was labeled FRET pair maleimide‐Atto 550 647N, alongside single‐stranded methyltetrazine‐modified oligonucleotide. experiments construct revealed structural transitions consistent previous studies, validating approach. Fluorescence measurements confirmed proximity pairs N‐terminally closed state experimental setup. This integrative method provides powerful tool complex dynamics beyond limitations traditional tweezers.

Language: Английский

Citations

0
Noncanonical Amino Acid Tools and Their Application to Membrane Protein Studies DOI Creative Commons

Chiara De Faveri,

Jordan M. Mattheisen, Thomas P. Sakmar

et al.

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(22), P. 12498 - 12550

Published: Nov. 7, 2024

Methods rooted in chemical biology have contributed significantly to studies of integral membrane proteins. One recent key approach has been the application genetic code expansion (GCE), which enables site-specific incorporation noncanonical amino acids (ncAAs) with defined properties into Efficient GCE is challenging, especially for proteins, specialized biogenesis and cell trafficking machinery tend be expressed at low levels membranes. Many eukaryotic proteins cannot functionally

Language: Английский

Citations

2
GPCR-targeted PROTACs in Cancer Therapeutics DOI Creative Commons
Victoria R. Saca, Thomas Huber, Thomas P. Sakmar

et al.

Molecular Pharmacology, Journal Year: 2024, Volume and Issue: 107(2), P. 100013 - 100013

Published: Dec. 12, 2024

G protein-coupled receptors (GPCRs) comprise a family of heptahelical membrane proteins that mediate intracellular and intercellular transmembrane signaling. Defects in GPCR signaling pathways are implicated the pathophysiology many diseases, including cardiovascular disease, endocrinopathies, immune disorders, cancer. Although GPCRs attractive drug targets, only small number Food Drug Administration-approved anticancer therapeutics target GPCRs. Targeted protein degradation (TPD) technology allows for direct modulation cellular expression level interest. TPD methods such as proteolysis-targeting chimeras (PROTACs) use ubiquitin-proteasome system to degrade interest selectively. PROTAC has not been widely applied other proteins, there is evidence PROTACs or could be GPCRome. Current show feasibility using GPCRs; however, mechanism some these uncertain. Additional studies aimed at elucidating with necessary. Discovery new allosteric molecule binders will required development intracellularly oriented PROTACs. Promising early results targeted suggest discovery platforms useful developing targeting pathological SIGNIFICANCE STATEMENT: Aberrant can contribute generally highly individual currently undrugged traditional approaches. technologies, chimeras, provide approach previously undruggable relevant molecular

Language: Английский

Citations

1
One-Pot Dual Protein Labelling for Simultaneous Mechanical and Fluorescent Readouts in Optical Tweezers DOI Creative Commons
Laura‐Marie Silbermann, Maximilian Fottner, Nora Migdad

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 19, 2024

Abstract Optical tweezers are widely used in the study of biological macromolecules but limited by their one-directional probing capability, potentially missing critical conformational changes. Combining fluorescence microscopy with optical tweezers, employing Förster resonance energy transfer (FRET) pairs, addresses this issue. Moreover, attaching one FRET probe to a tethered protein and other solution allows precise localisation interaction sites, while mechanical properties. When integrating orthogonal conjugation methods needed enable simultaneous, site-specific attachment fluorophores DNA handles, commonly apply force molecules interest. In study, we utilized commercially available reagents for dual labelling homodimeric heat shock 90 (Hsp90) using thiol-maleimide inverse electron demand Diels–Alder cycloaddition (IEDDAC) bioorthogonal reactions. one-pot approach, Hsp90 modified cysteine mutation non-canonical amino acid cyclopropene-L-lysine (CpK) was labelled pair maleimide-Atto550 maleimide-Atto647N, alongside single- stranded methyltetrazine-modified oligonucleotide. experiments construct revealed structural transitions consistent previous studies, validating approach. Fluorescence measurements confirmed proximity pairs N-terminally closed state experimental setup. This integrative method provides powerful tool dynamics interactions beyond limitations traditional tweezers. Statement The developed combines enhancing single-molecule studies overcoming one- directional probing. Utilizing two labelling, single-stranded oligonucleotides. Validated comparison published changes, unfolding signatures, distances, approach explore interactions.

Language: Английский

Citations

0
Development of a Class A/B Hybrid GPCR System for the Proximity-Assisted Screening of GPCR Ligands DOI

Monika T. Gnatzy,

S. Hartmann, Felix Hausch

et al.

ACS Chemical Biology, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 16, 2024

Class A G protein-coupled receptors (GPCRs) are key mediators in numerous signaling pathways and important drug targets for several diseases. major shortcoming GPCR ligand screening is the detection limit weak binding molecules, which especially critical poorly druggable GPCRs. Here, we present a proximity-based system class GPCRs, adopts natural two-step activation mechanism of B In this approach, A/B chimeras with extracellular domain receptor CRF1R grafted to transmembrane target stimulated hybrid ligands. These ligands contain high-affinity peptide derived from CRF, recruits engineered GPCR, dramatically increasing local concentration test substances. We exemplified method neurotensin 1 (NTR1) endothelin (ETB), two pulmonary arterial hypertension or psychological disorders neurodegenerative observed >20× activity enhancement by directed proximity enabling weakly activating sequences that would have otherwise remained undetected. Our approach allows probe membrane living cells may be useful GPCRs it has been difficult generate small drug-like molecules.

Language: Английский

Citations

0
Genetic Code Expansion: Recent Developments and Emerging Applications DOI Creative Commons
Yujia Huang, Pan Zhang, Haoyu Wang

et al.

Chemical Reviews, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 31, 2024

The concept of genetic code expansion (GCE) has revolutionized the field chemical and synthetic biology, enabling site-specific incorporation noncanonical amino acids (ncAAs) into proteins, thus opening new avenues in research applications across biology medicine. In this review, we cover principles GCE, including optimization aminoacyl-tRNA synthetase (aaRS)/tRNA system advancements translation engineering. Notable developments include refinement aaRS/tRNA pairs, enhancements screening methods, biosynthesis acids. GCE technology span from where it facilitates gene expression regulation protein engineering, to medicine, with promising approaches drug development, vaccine production, editing. review concludes a perspective on future underscoring its potential further expand toolkit Through comprehensive aim provide detailed overview current state technology, challenges, opportunities, frontier represents for novel biological therapeutic applications.

Language: Английский

Citations

0
Biophysical Insights into Drug Discovery: Leveraging Phase Transitions and Protein Behavior for Therapeutic Innovation DOI
Afam Uzorka,

David Kibirige,

David Makumbi

et al.

Biophysical Reviews and Letters, Journal Year: 2023, Volume and Issue: 18(03), P. 171 - 176

Published: Sept. 1, 2023

In the search for novel treatments various diseases, nexus of biophysics and drug discovery represents a dynamic transformational paradigm. A new age in pharmaceutical research has begun thanks to its thorough grasp structural physical characteristics biological molecules. This study explores how advances biophysical approaches, including allosteric modulation, intrinsically disordered proteins (IDPs), protein phase transitions, have transformed development process. Protein supported by principles, provided crucial insights into disorders like ALS Alzheimer’s, opening up avenues therapeutic intervention. Targeting IDPs their role liquid–liquid separation produced creative approaches diseases that were formerly thought be resistant therapy. order reduce complexity complicated concept made possible understanding, offers precise selective approach medication creation. review highlights significant influence on therapeutics drugs. Biophysics is advancing discipline toward creation more precise, efficient, tailored medications illuminating properties proteins, complexities dynamics interactions. Biophysical are poised alter healthcare bringing together several fields sustained innovation, giving people dealing with variety ailments fresh hope.

Language: Английский

Citations

1