Detection of Transport Intermediates in the Peptidoglycan Flippase MurJ Identifies Residues Essential for Conformational Cycling.

Journal of the American Chemical Society
Frederick A RubinoDaniel Kahne

Abstract

Bacterial cell wall synthesis is an essential process in bacteria and one of the best targets for antibiotics. A critical step on this pathway is the export of the lipid-linked cell wall monomer, Lipid II, by its transporter MurJ. The mechanism by which MurJ mediates the transbilayer movement of Lipid II is not understood because intermediate states of this process have not been observed. Here we demonstrate a method to capture and detect interactions between MurJ and its substrate Lipid II by photo-cross-linking and subsequent biotin-tagging. We show that this method can be used to covalently capture intermediate transport states of Lipid II on MurJ in living cells. Using this strategy we probed several lethal arginine mutants and found that they retain appreciable substrate-binding ability despite being defective in Lipid II transport. We propose that Lipid II binding to these residues during transport induces a conformational change in MurJ required to proceed through the Lipid II transport cycle. The methods described to detect intermediate transport states of MurJ will be useful for characterizing mechanisms of inhibitors.

References

Aug 3, 2002·Proceedings of the National Academy of Sciences of the United States of America·Jason W ChinPeter G Schultz
Feb 27, 2003·European Journal of Biochemistry·Rikki N HvorupMilton H Saier
Oct 4, 2008·Proceedings of the National Academy of Sciences of the United States of America·Natividad Ruiz
Jun 6, 2012·Annual Review of Biochemistry·Andrew L LoveringNatalie C J Strynadka
Sep 17, 2014·Journal of Bacteriology·Emily K ButlerNatividad Ruiz
Oct 8, 2014·Journal of the American Chemical Society·Yuan QiaoSuzanne Walker
Mar 30, 2016·Annual Review of Biochemistry·David Drew, Olga Boudker
May 27, 2016·Angewandte Chemie·Sourav SarkarMarcos M Pires
Dec 27, 2016·Nature Structural & Molecular Biology·Alvin C Y KukSeok-Yong Lee
May 30, 2017·Nature Chemical Biology·Yuan QiaoDaniel Kahne
Nov 29, 2017·Journal of the American Chemical Society·Michael A WelshSuzanne Walker
Feb 21, 2018·Nature Chemistry·Jani Reddy BollaCarol V Robinson
Mar 21, 2018·Journal of the American Chemical Society·Frederick A RubinoDaniel E Kahne
Jun 13, 2018·Proceedings of the National Academy of Sciences of the United States of America·Sanduo ZhengAndrew C Kruse
Jun 20, 2018·Proceedings of the National Academy of Sciences of the United States of America·Emel FiciciJosé D Faraldo-Gómez
Nov 30, 2018·The Journal of Biological Chemistry·Sujeet KumarNatividad Ruiz
Apr 17, 2019·Nature Communications·Alvin C Y KukSeok-Yong Lee

❮ Previous
Next ❯

Citations

May 20, 2020·The Journal of Biological Chemistry·Chris WhitfieldSteven D Kelly
Dec 19, 2020·FEMS Microbiology Reviews·Tong SuLok-To Sham
Aug 22, 2020·Cell Chemical Biology·Ashley R BrownCatherine L Grimes
Sep 22, 2020·Chemical Reviews·Emily LundstedtNatividad Ruiz

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.