Peptide Inhibitor Assay for Allocating Functionally Important Accessible Sites Throughout a Protein Chain: Restriction Endonuclease EcoRI as a Model Protein System DOI Creative Commons
Joji M. Otaki

BioTech, Год журнала: 2024, Номер 14(1), С. 1 - 1

Опубликована: Дек. 30, 2024

Functionally important amino acid sequences in proteins are often located at multiple sites. Three-dimensional structural analysis and site-directed mutagenesis may be performed to allocate functional sites for understanding structure‒function relationships developing novel inhibitory drugs. However, such methods too demanding comprehensively cover potential throughout a protein chain. Here, peptide inhibitor assay (PIA) was devised functionally accessible proteins. This simple method presumes that protein‒ligand interactions, intramolecular dimerization interactions can partially inhibited by high concentrations of competitive “endogenous” peptides the interest. Focusing on restriction endonuclease EcoRI as model system, many endogenous (6mer-14mer) were synthesized, covering entire Some them highly inhibitory, but interestingly, nine most effective outside active sites, with exception one. Relatively long aromatic residues (F, H, W, Y) corresponding secondary structures generally effective. Because synthetic flexible enough change length residues, this useful quickly drugs or epitopes neutralizing antibodies.

Язык: Английский

Molecular/antigenic mimicry and immunological cross-reactivity explains SARS-CoV-2-induced autoimmunity DOI
Yekbun Adıgüzel, Dimitrios P. Bogdanos, Yehuda Shoenfeld

и другие.

Autoimmunity Reviews, Год журнала: 2025, Номер unknown, С. 103811 - 103811

Опубликована: Апрель 1, 2025

Язык: Английский

Процитировано

0
Antibody-Mediated Protein Knockdown Reveals Distal-less Functions for Eyespots and Parafocal Elements in Butterfly Wing Color Pattern Development DOI Creative Commons

Yugo Nakazato,

Joji M. Otaki

Cells, Год журнала: 2024, Номер 13(17), С. 1476 - 1476

Опубликована: Сен. 2, 2024

One of the important genes for eyespot development in butterfly wings is Distal-less. Its function has been evaluated via several methods, including CRISPR/Cas9 genome editing. However, functional inhibition may be performed at right time place using a different method. Here, we used novel protein delivery method pupal wing tissues vivo to inactivate target protein, Distal-less, with polyclonal anti-Distal-less antibody blue pansy Junonia orithya. We first demonstrated that various antibodies were delivered epithelial cells this species. Treatment reduced size, confirming positive role Distal-less development. The treatment eliminated or deformed parafocal element, suggesting element. This result also suggested integrity an and its corresponding element as border symmetry system. Taken together, these findings demonstrate antibody-mediated knockdown useful tool assays proteins, such expressed tissues, functions eyespots elements color pattern

Язык: Английский

Процитировано

3
Peptide Inhibitor Assay for Allocating Functionally Important Accessible Sites Throughout a Protein Chain: Restriction Endonuclease EcoRI as a Model Protein System DOI Creative Commons
Joji M. Otaki

BioTech, Год журнала: 2024, Номер 14(1), С. 1 - 1

Опубликована: Дек. 30, 2024

Functionally important amino acid sequences in proteins are often located at multiple sites. Three-dimensional structural analysis and site-directed mutagenesis may be performed to allocate functional sites for understanding structure‒function relationships developing novel inhibitory drugs. However, such methods too demanding comprehensively cover potential throughout a protein chain. Here, peptide inhibitor assay (PIA) was devised functionally accessible proteins. This simple method presumes that protein‒ligand interactions, intramolecular dimerization interactions can partially inhibited by high concentrations of competitive “endogenous” peptides the interest. Focusing on restriction endonuclease EcoRI as model system, many endogenous (6mer-14mer) were synthesized, covering entire Some them highly inhibitory, but interestingly, nine most effective outside active sites, with exception one. Relatively long aromatic residues (F, H, W, Y) corresponding secondary structures generally effective. Because synthetic flexible enough change length residues, this useful quickly drugs or epitopes neutralizing antibodies.

Язык: Английский

Процитировано

0