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The origins of specificity in the microcin-processing protease TldD/E
DOI:
10.1016/j.str.2017.08.006
Authors:
Dmitry
Ghilarov
(Skolkovo Institute of Science and Technology)
,
Marina
Serebryakova
(Institute of Gene Biology of the Russian Academy of Sciences; Lomonosov Moscow State University)
,
Clare E. M.
Stevenson
(John Innes Centre)
,
Stephen J.
Hearnshaw
(John Innes Centre)
,
Dmitry
Volkov
(Lomonosov Moscow State University)
,
Anthony
Maxwell
(John Innes Centre)
,
David M.
Lawson
(John Innes Centre)
,
Konstantin
Severinov
(Skolkovo Institute of Science and Technology; Bionano Institute, Peter the Great Saint Petersburg State Polytechnical University; The State University of New Jersey)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Structure
State:
Published (Approved)
Published:
September 2017
Diamond Proposal Number(s):
9475

Abstract: TldD and TldE proteins are involved in the biosynthesis of microcin B17 (MccB17), an Escherichia coli thiazole/oxazole-modified peptide toxin targeting DNA gyrase. Using a combination of biochemical and crystallographic methods we show that E. coli TldD and TldE interact to form a heterodimeric metalloprotease. TldD/E cleaves the N-terminal leader sequence from the modified MccB17 precursor peptide, to yield mature antibiotic, while it has no effect on the unmodified peptide. Both proteins are essential for the activity; however, only the TldD subunit forms a novel metal-containing active site within the hollow core of the heterodimer. Peptide substrates are bound in a sequence-independent manner through β sheet interactions with TldD and are likely cleaved via a thermolysin-type mechanism. We suggest that TldD/E acts as a “molecular pencil sharpener”: unfolded polypeptides are fed through a narrow channel into the active site and processively truncated through the cleavage of short peptides from the N-terminal end.
Journal Keywords: microcin B17; DNA gyrase; metalloprotease; CcdAB; X-ray crystallography; toxin-antitoxin; ribosomally synthesized modified peptides; RiPP; peptidase
Diamond Keywords: Bacteria
Subject Areas:
Biology and Bio-materials,
Chemistry,
Medicine
Instruments:
I02-Macromolecular Crystallography
,
I03-Macromolecular Crystallography
,
I04-1-Macromolecular Crystallography (fixed wavelength)
Added On:
23/09/2017 13:27
Documents:
1-s2.0-S0969212617302599-main.pdf
Discipline Tags:
Pathogens
Antibiotic Resistance
Infectious Diseases
Health & Wellbeing
Biochemistry
Genetics
Chemistry
Structural biology
Life Sciences & Biotech
Technical Tags:
Diffraction
Macromolecular Crystallography (MX)