scholarly journals Gated Proton Release during Radical Transfer at the Subunit Interface of Ribonucleotide Reductase

2020 ◽  
Author(s):  
Chang Cui ◽  
Brandon L. Greene ◽  
Gyunghoon Kang ◽  
Catherine L. Drennan ◽  
JoAnne Stubbe ◽  
...  
Keyword(s):  
Ribonucleotide Reductase ◽  
Proton Release ◽  
Subunit Interface ◽  
Radical Transfer

scholarly journals Glutamate 52-β at the α/β subunit interface ofEscherichia coliclass Ia ribonucleotide reductase is essential for conformational gating of radical transfer

2017 ◽  
Vol 292 (22) ◽  
pp. 9229-9239 ◽  
Author(s):  
Qinghui Lin ◽  
Mackenzie J. Parker ◽  
Alexander T. Taguchi ◽  
Kanchana Ravichandran ◽  
Albert Kim ◽  
...  
Keyword(s):  
Ribonucleotide Reductase ◽  
Β Subunit ◽  
Subunit Interface ◽  
Radical Transfer ◽  
Conformational Gating

scholarly journals Radical transfer in E. coli ribonucleotide reductase: a NH2Y731/R411A-α mutant unmasks a new conformation of the pathway residue 731

2016 ◽  
Vol 7 (3) ◽  
pp. 2170-2178 ◽  
Author(s):  
Müge Kasanmascheff ◽  
Wankyu Lee ◽  
Thomas U. Nick ◽  
JoAnne Stubbe ◽  
Marina Bennati
Keyword(s):  
Long Range ◽  
Ribonucleotide Reductase ◽  
E Coli ◽  
Subunit Interface ◽  
Radical Transfer

A new conformation of theE. coliRNR pathway residue 731 was trapped during long-range radical transfer across the αβ subunit interface.

scholarly journals In Vivo Assay for Low-Activity Mutant Forms of Escherichia coli Ribonucleotide Reductase

2003 ◽  
Vol 185 (4) ◽  
pp. 1167-1173 ◽  
Author(s):  
Monica Ekberg ◽  
Pernilla Birgander ◽  
Britt-Marie Sjöberg
Keyword(s):  
Escherichia Coli ◽  
Ribonucleotide Reductase ◽  
Active Site ◽  
Site Directed Mutagenesis ◽  
In Vitro Assays ◽  
Tyrosyl Radical ◽  
In Vivo Assay ◽  
Radical Transfer

ABSTRACT Ribonucleotide reductase (RNR) catalyzes the essential production of deoxyribonucleotides in all living cells. In this study we have established a sensitive in vivo assay to study the activity of RNR in aerobic Escherichia coli cells. The method is based on the complementation of a chromosomally encoded nonfunctional RNR with plasmid-encoded RNR. This assay can be used to determine in vivo activity of RNR mutants with activities beyond the detection limits of traditional in vitro assays. E. coli RNR is composed of two homodimeric proteins, R1 and R2. The R2 protein contains a stable tyrosyl radical essential for the catalysis that takes place at the R1 active site. The three-dimensional structures of both proteins, phylogenetic studies, and site-directed mutagenesis experiments show that the radical is transferred from the R2 protein to the active site in the R1 protein via a radical transfer pathway composed of at least nine conserved amino acid residues. Using the new assay we determined the in vivo activity of mutants affecting the radical transfer pathway in RNR and identified some residual radical transfer activity in two mutant R2 constructs (D237N and W48Y) that had previously been classified as negative for enzyme activity. In addition, we show that the R2 mutant Y356W is completely inactive, in sharp contrast to what has previously been observed for the corresponding mutation in the mouse R2 enzyme.

scholarly journals Conformationally Dynamic Radical Transfer within Ribonucleotide Reductase

2017 ◽  
Vol 139 (46) ◽  
pp. 16657-16665 ◽  
Author(s):  
Brandon L. Greene ◽  
Alexander T. Taguchi ◽  
JoAnne Stubbe ◽  
Daniel G. Nocera
Keyword(s):  
Ribonucleotide Reductase ◽  
Radical Transfer

Spectroscopic Evidence for a H Bond Network at Y356 Located at the Subunit Interface of Active E. coli Ribonucleotide Reductase

Biochemistry ◽  
2017 ◽  
Vol 56 (28) ◽  
pp. 3647-3656 ◽  
Author(s):  
Thomas U. Nick ◽  
Kanchana R. Ravichandran ◽  
JoAnne Stubbe ◽  
Müge Kasanmascheff ◽  
Marina Bennati
Keyword(s):  
Ribonucleotide Reductase ◽  
Spectroscopic Evidence ◽  
E Coli ◽  
Subunit Interface ◽  
Bond Network

scholarly journals Preserved Catalytic Activity in an Engineered Ribonucleotide Reductase R2 Protein with a Nonphysiological Radical Transfer Pathway

1998 ◽  
Vol 273 (33) ◽  
pp. 21003-21008 ◽  
Author(s):  
Monica Ekberg ◽  
Stephan Pötsch ◽  
Elin Sandin ◽  
Marjolein Thunnissen ◽  
Pär Nordlund ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Ribonucleotide Reductase ◽  
Radical Transfer ◽  
Ribonucleotide Reductase R2
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