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1.
Chembiochem ; 22(21): 3037-3041, 2021 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-34018291

RESUMO

The major capsid protein VP1 of JC Polyomavirus assembles into pentamers that serve as a model for studying viral entry of this potentially severe human pathogen. Previously, labeling of viral proteins utilized large fusion proteins or non-specific amine- or cysteine-functionalization with fluorescent dyes. Imaging of these sterically hindered fusion proteins or heterogeneously labeled virions limits reproducibility and could prevent the detection of subtle trafficking phenomena. Here we advance the π-clamp-mediated cysteine conjugation for site-selective fluorescent labeling of VP1-pentamers. We demonstrate a one-step synthesis of a probe consisting of a bio-orthogonal click chemistry handle bridged to a perfluoro-biphenyl π-clamp reactive electrophile by a polyethylene glycol linker. We expand the scope of the π-clamp conjugation by demonstrating selective labeling of an internal, surface exposed loop in VP1. Thus, the π-clamp conjugation offers a general method to selectively bioconjugate tags-of-interest to viral proteins without impeding their ability to bind and enter cells.


Assuntos
Proteínas do Capsídeo/metabolismo , Cisteína/metabolismo , Vírus JC/metabolismo , Bibliotecas de Moléculas Pequenas/metabolismo , Proteínas do Capsídeo/química , Cisteína/química , Vírus JC/química , Modelos Moleculares , Estrutura Molecular , Bibliotecas de Moléculas Pequenas/química
2.
J Control Release ; 326: 106-119, 2020 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-32569705

RESUMO

The gut microbiome is a promising target for the development of GI tract therapies, yet it has been under-exploited due, in part, to a lack of tools to control and manipulate complex microbial communities. To date, the most common approach in harnessing bacteria for therapeutic purposes has been to deliver ex vivo engineered bacteria-effectively taking a bacterial cell therapy-based approach. An alternative approach involves taking advantage of the rich microbial ecosystem in the gut by genetically modifying the microbiome in situ through the use of engineered bacteriophages-akin to human gene therapies delivered by viral vectors. In this review, we present the challenges and opportunities associated with engineering bacteriophages to control and manipulate the gut microbiome.


Assuntos
Bacteriófagos , Microbioma Gastrointestinal , Microbiota , Bactérias , Humanos
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