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

2020 ◽  
Vol 56 (82) ◽  
pp. 12319-12322
Author(s):  
Jerrin Thomas George ◽  
Seergazhi G. Srivatsan
Keyword(s):  
Fluorescent Nucleotides ◽  
Enzyme Recognition

A terminal uridylyl transferase site-specifically labels RNA with microenvironment-sensitive fluorescent nucleotides, which in turn provide direct read-outs to probe the enzyme recognition.

scholarly journals Serpin reactive center loop mobility is required for inhibitor function but not for enzyme recognition.

1994 ◽  
Vol 269 (44) ◽  
pp. 27657-27662 ◽  
Author(s):  
D A Lawrence ◽  
S T Olson ◽  
S Palaniappan ◽  
D Ginsburg
Keyword(s):  
Reactive Center Loop ◽  
Reactive Center ◽  
Enzyme Recognition

scholarly journals Activation mechanism of ATP-sensitive K+ channels explored with real-time nucleotide binding

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Michael Puljung ◽  
Natascia Vedovato ◽  
Samuel Usher ◽  
Frances Ashcroft
Keyword(s):  
Conformational Change ◽  
Ionic Currents ◽  
Nucleotide Binding ◽  
Activation Mechanism ◽  
Time Resolved ◽  
K Channels ◽  
Novel Approach ◽  
Binding Event ◽  
Fluorescent Nucleotides

The response of ATP-sensitive K+ channels (KATP) to cellular metabolism is coordinated by three classes of nucleotide binding site (NBS). We used a novel approach involving labeling of intact channels in a native, membrane environment with a non-canonical fluorescent amino acid and measurement (using FRET with fluorescent nucleotides) of steady-state and time-resolved nucleotide binding to dissect the role of NBS2 of the accessory SUR1 subunit of KATP in channel gating. Binding to NBS2 was Mg2+-independent, but Mg2+ was required to trigger a conformational change in SUR1. Mutation of a lysine (K1384A) in NBS2 that coordinates bound nucleotides increased the EC50 for trinitrophenyl-ADP binding to NBS2, but only in the presence of Mg2+, indicating that this mutation disrupts the ligand-induced conformational change. Comparison of nucleotide-binding with ionic currents suggests a model in which each nucleotide binding event to NBS2 of SUR1 is independent and promotes KATP activation by the same amount.

Distribution of Restriction Enzyme Recognition Sequences on Broad Host Range Plasmid RP4: Molecular and Evolutionary Implications

1996 ◽  
Vol 258 (3) ◽  
pp. 447-456 ◽  
Author(s):  
Brian M. Wilkins ◽  
Paul M. Chilley ◽  
Angela T. Thomas ◽  
Michael J. Pocklington
Keyword(s):  
Host Range ◽  
Restriction Enzyme ◽  
Broad Host Range ◽  
Broad Host Range Plasmid ◽  
Enzyme Recognition
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