Activatable Photoacoustic Probe for Imaging Infection: Gold Nanorod Dissociation In Vivo Reports Bacterial Protease Activity DOI
Maurice Retout, Victor Lepeintre, Lubna Amer

et al.

ACS Nano, Journal Year: 2025, Volume and Issue: unknown

Published: March 21, 2025

We present a strategy for constructing activatable photoacoustic imaging (PAI) probes in vivo enzyme activity measurements, based on dissociation previously applied to vitro sensing. Unlike conventional nanoparticle aggregation strategies, minimizes false positives and functions effectively complex biological environments. Overcoming the challenge of dissociating nanostructure aggregates, which arises from strong van der Waals forces at short distances, we demonstrate controlled assembly citrate-capped gold nanorods (AuNRs-citrate) using diarginine peptide additive thiolated polyethylene glycol (HS-PEG-OMe), respectively. This mechanism enables precise control optical (PA) properties AuNRs both settings. Building these findings, engineered an enzyme-sensitive PAI probe (AuNRs@RgpB) composed AuNR assemblies PEG-peptide conjugate with protease-specific cleavage sequence. The detects Arg-specific gingipain (RgpB), protease expressed by Porphyromonas gingivalis associated periodontal disease Alzheimer's disease. Proteolytic sequence triggers dissociation, resulting enhanced PA signal output. was designed be injected intrathecally preclinical trials image gingipains investigate value inhibitors developed probe's performance characterized UV-vis spectroscopy PAI, achieving detection limits 5 20 nM, In studies involved intracranial injection AuNRs@RgpB into RgpB-containing murine models, monitoring over time. RgpB produced four-fold increase within 2 h, while P. gingivalis-infected mice showed similar enhancement. Specificity confirmed negligible responses Fusobacterium nucleatum, non-RgpB-producing bacterium. Additionally, system demonstrated utility drug development successfully inhibition (leupeptin KYT-1) models. Beyond its immediate application detection, this modular approach plasmonic-based sensing holds significant potential detecting other proteases, advancing nanotechnology protease-targeted diagnostics.

Language: Английский

Spacer Matters: All-Peptide-Based Ligand for Promoting Interfacial Proteolysis and Plasmonic Coupling DOI
Zhicheng Jin, Chuxuan Ling, Yi Li

et al.

Nano Letters, Journal Year: 2022, Volume and Issue: 22(22), P. 8932 - 8940

Published: Nov. 8, 2022

Plasmonic coupling via nanoparticle assembly is a popular signal-generation method in bioanalytical sensors. Here, we customized an all-peptide-based ligand that carries anchoring group, polyproline spacer, biomolecular recognition, and zwitterionic domains for functionalizing gold nanoparticles (AuNPs) as colorimetric enzyme sensor. Our results underscore the importance of module, which enables SARS-CoV-2 main protease (Mpro) to recognize peptidic on nanosurfaces subsequent plasmonic Coulombic interactions. AuNP aggregation favored by lowered surface potential due enzymatic unveiling module. Therefore, this system provides naked-eye measure Mpro. No proteolysis occurs AuNPs modified with control lacking spacer domain. Overall, does not require complex molecular conjugations hence offers simple promising route sensing other proteases.

Language: Английский

Citations

21
Peptidic Sulfhydryl for Interfacing Nanocrystals and Subsequent Sensing of SARS-CoV-2 Protease DOI
Zhicheng Jin, Justin Yeung, Jiajing Zhou

et al.

Chemistry of Materials, Journal Year: 2022, Volume and Issue: 34(3), P. 1259 - 1268

Published: Jan. 18, 2022

There is a need for surveillance of COVID-19 to identify individuals infected with SARS-CoV-2 coronavirus. Although specific, nucleic acid testing has limitations in terms point-of-care testing. One potential alternative the nonstructural protease (nsp5, also known as Mpro/3CLpro) implicated viral replication but not incorporated into virions. Here, we report divalent substrate novel design, (Cys)2–(AA)x–(Asp)3, interface gold colloids specific presence Mpro leading rapid and colorimetric readout. Citrate- tris(2-carboxyethyl)phosphine (TCEP)-AuNPs were identified best reporter out 17 ligated nanoparticles. Furthermore, empirically determined effects varying cysteine valence biological media on sensor specificity sensitivity. The peptide was Mpro, that is, there no response when tested other proteins or enzymes. detection limits Tris buffer exhaled breath matrices are 12.2 18.9 nM, respectively, which comparable reported methods (i.e., at low nanomolar concentrations) yet visual These results from our work would provide informative rationales design practical noninvasive diagnostic testing─the proteases biofluids validated.

Language: Английский

Citations

19
A Protease-Responsive Polymer/Peptide Conjugate and Reversible Assembly of Silver Clusters for the Detection of Porphyromonas gingivalis Enzymatic Activity DOI
Maurice Retout, Lubna Amer, Wonjun Yim

et al.

ACS Nano, Journal Year: 2023, Volume and Issue: 17(17), P. 17308 - 17319

Published: Aug. 21, 2023

We report the reversible aggregation of silver nanoparticle (AgNP) assemblies using combination a cationic arginine-based peptide and sulfur-capped polyethylene glycol (PEG). The formation dissociation aggregates were studied by optical methods electron microscopy. clusters depends on sequence PEG size. A molecular weight 1 kDa for was optimal dissociation. most important feature this method is that it can operate in complex biofluids such as plasma, saliva, bile, urine, cell media, or even seawater without significant decrease performance. Moreover, peptide-particle are highly stable do not degrade (or express loss signal upon dissociation) when dried resolubilized, frozen thawed, left daylight month. Importantly, capacity be reduced via conjugation peptide-cleavable substrate. restored presence an enzyme. Based these findings, we designed PEG-peptide hybrid molecule specific to Porphyromonas gingivalis protease RgpB. Our motivation bacterium key pathogen periodontitis, RgpB activity has been correlated with chronic diseases including Alzheimer's disease. limit detection 100 pM vitro. This system used measure gingival crevicular fluid (GCF) samples rate 40% 0% false negatives versus PCR P. (n = 37). nanoparticles allows development convenient sensing independently media composition.

Language: Английский

Citations

12
Determination of SARS-CoV-2 Main Protease (Mpro) Activity Based on Electrooxidation of Tyrosine Residue of a Model Peptide DOI
Tatiana A. Filippova, Rami A. Masamrekh,

Tatiana Farafonova

et al.

Biochemistry (Moscow), Journal Year: 2025, Volume and Issue: 90(1), P. 120 - 131

Published: Jan. 1, 2025

Language: Английский

Citations

0
Activatable Photoacoustic Probe for Imaging Infection: Gold Nanorod Dissociation In Vivo Reports Bacterial Protease Activity DOI
Maurice Retout, Victor Lepeintre, Lubna Amer

et al.

ACS Nano, Journal Year: 2025, Volume and Issue: unknown

Published: March 21, 2025

We present a strategy for constructing activatable photoacoustic imaging (PAI) probes in vivo enzyme activity measurements, based on dissociation previously applied to vitro sensing. Unlike conventional nanoparticle aggregation strategies, minimizes false positives and functions effectively complex biological environments. Overcoming the challenge of dissociating nanostructure aggregates, which arises from strong van der Waals forces at short distances, we demonstrate controlled assembly citrate-capped gold nanorods (AuNRs-citrate) using diarginine peptide additive thiolated polyethylene glycol (HS-PEG-OMe), respectively. This mechanism enables precise control optical (PA) properties AuNRs both settings. Building these findings, engineered an enzyme-sensitive PAI probe (AuNRs@RgpB) composed AuNR assemblies PEG-peptide conjugate with protease-specific cleavage sequence. The detects Arg-specific gingipain (RgpB), protease expressed by Porphyromonas gingivalis associated periodontal disease Alzheimer's disease. Proteolytic sequence triggers dissociation, resulting enhanced PA signal output. was designed be injected intrathecally preclinical trials image gingipains investigate value inhibitors developed probe's performance characterized UV-vis spectroscopy PAI, achieving detection limits 5 20 nM, In studies involved intracranial injection AuNRs@RgpB into RgpB-containing murine models, monitoring over time. RgpB produced four-fold increase within 2 h, while P. gingivalis-infected mice showed similar enhancement. Specificity confirmed negligible responses Fusobacterium nucleatum, non-RgpB-producing bacterium. Additionally, system demonstrated utility drug development successfully inhibition (leupeptin KYT-1) models. Beyond its immediate application detection, this modular approach plasmonic-based sensing holds significant potential detecting other proteases, advancing nanotechnology protease-targeted diagnostics.

Language: Английский

Citations

0