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Responsive fluorescent nucleotides serve as efficient substrates to probe terminal uridylyl transferase.

ABSTRACT: We repurposed a terminal uridylyl transferase enzyme to site-specifically label RNA with microenvironment sensing fluorescent nucleotide mimics, which in turn provided direct read-outs to estimate the binding affinities of the enzyme to RNA and nucleotide substrates. This enzyme-probe system provides insights into the catalytic cycle, and can facilitate the development of discovery platforms to identify robust enzyme inhibitors.

SUBMITTER: George JT

PROVIDER: S-EPMC7611084 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Responsive fluorescent nucleotides serve as efficient substrates to probe terminal uridylyl transferase.

George Jerrin Thomas JT Srivatsan Seergazhi G SG

Chemical communications (Cambridge, England) 20200917 82

We repurposed a terminal uridylyl transferase enzyme to site-specifically label RNA with microenvironment sensing fluorescent nucleotide mimics, which in turn provided direct read-outs to estimate the binding affinities of the enzyme to RNA and nucleotide substrates. This enzyme-probe system provides insights into the catalytic cycle, and can facilitate the development of discovery platforms to identify robust enzyme inhibitors. ...[more]

PMID: 32939524

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