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Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
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The two-state receptor model explains a drug's interaction with receptors, such as G protein-coupled receptors and ligand-gated ion channels, to induce or inhibit a biological response. When no natural ligands are present, a receptor exists in an equilibrium of inactive (Ri) and active (Ra) conformations. The inactive form does not produce a response, while the active form generates a basal effect known as constitutive activity.
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Select azo compounds post-translationally modulate HTRA1 abundance and activity potentially through interactions at

John D Hulleman1, Seungje Jeon2, Sofia Bali3,4

  • 1Department of Ophthalmology and Visual Neurosciences, University of Minnesota, 2001 6 St. SE, Minneapolis, Minnesota, 55455, United States.

Biorxiv : the Preprint Server for Biology
|June 4, 2025
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High-temperature requirement protein A1 (HTRA1) is crucial for eye health and linked to age-related macular degeneration (AMD). Researchers screened compounds and found Chicago Sky Blue 6B enhances HTRA1 levels, offering potential therapeutic avenues for AMD.

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Area of Science:

  • Molecular Biology
  • Neuroscience
  • Ophthalmology

Background:

  • High-temperature requirement protein A1 (HTRA1) is a serine protease implicated in neurodegenerative diseases, including age-related macular degeneration (AMD).
  • Genetic studies link the 10q26 locus containing HTRA1 to AMD susceptibility, with risk alleles potentially causing age-related defects in retinal HTRA1 synthesis.

Purpose of the Study:

  • To identify small molecules that enhance HTRA1 transcription or protein levels using a high-throughput screening approach.
  • To explore potential therapeutic strategies for AMD by targeting HTRA1.

Main Methods:

  • CRISPR/Sp.Cas9 gene editing was used to tag HTRA1 with a HiBiT peptide in ARPE-19 cells.
  • A library of 1920 compounds was screened for their effect on HTRA1 secretion and intracellular levels.
  • Counter-screening, HiBiT blotting, serine hydrolase activity-based protein profiling (SH-ABPP), and computational modeling were employed.

Main Results:

  • Chicago Sky Blue 6B (CSB), an azo compound, significantly enhanced HTRA1 secretion (2.0-fold) and intracellular levels (1.7-fold) without altering transcription or enzymatic activity.
  • Congo Red, another azo compound, increased intracellular HTRA1 (3.6-fold) but impaired its enzymatic activity.
  • Computational modeling suggested CSB and Congo Red bind to HTRA1's trimer interface.

Conclusions:

  • Select azo dyes, including CSB, can serve as chemical probes to modulate HTRA1 levels.
  • These findings identify potential starting points for developing novel small molecule therapeutics targeting HTRA1 for AMD and other related conditions.