Identification of essential amino acid residues in the nisin dehydratase NisB

Frontiers in Microbiology
Rustem KhusainovO P Kuipers

Abstract

Nisin is a posttranslationally-modified antimicrobial peptide that has the ability to induce its own biosynthesis. Serines and threonines in the modifiable core peptide part of precursor nisin are dehydrated to dehydroalanines and dehydrobutyrines by the dehydratase NisB, and subsequently cysteines are coupled to the dehydroamino acids by the cyclase NisC. In this study, we applied extensive site-directed mutagenesis, together with direct binding studies, to investigate the molecular mechanism of the dehydratase NisB. We use a natural nisin-producing strain as a host to probe mutant-NisB functionality. Importantly, we are able to differentiate between intracellular and secreted fully dehydrated precursor nisin, enabling investigation of the NisB properties needed for the release of dehydrated precursor nisin to its devoted secretion system NisT. We report that single amino acid substitutions of conserved residues, i.e., R83A, R83M, and R87A result in incomplete dehydration of precursor nisin and prevention of secretion. Single point NisB mutants Y80F and H961A, result in a complete lack of dehydration of precursor nisin, but do not abrogate precursor nisin binding. The data indicate that residues Y80 and H961 are directly invol...Continue Reading

References

Oct 1, 1996·Applied and Environmental Microbiology·P G de RuyterW M de Vos
Jan 23, 2004·BMC Bioinformatics·Benjamin Schuster-BöcklerSven Rahmann
Jan 31, 2004·Science·Lili XieWilfred A van der Donk
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Nov 3, 2005·Journal of the American Chemical Society·Champak ChatterjeeWilfred A van der Donk
May 22, 2007·The Journal of Biological Chemistry·Bo Li, Wilfred A van der Donk
Oct 30, 2007·Cellular and Molecular Life Sciences : CMLS·J LubelskiO P Kuipers
Jul 16, 2008·Applied and Environmental Microbiology·H Bart van den Berg van SaparoeaArnold J M Driessen
Sep 16, 2008·Applied and Environmental Microbiology·Anneke KuipersGert N Moll
Nov 29, 2008·The Journal of Pharmacology and Experimental Therapeutics·Leon D KluskensMarijke Haas
Jul 22, 2009·The Journal of Biological Chemistry·Jacek LubelskiOscar P Kuipers
Aug 12, 2009·Journal of the American Chemical Society·M Violet LeeNeil L Kelleher
Feb 24, 2010·Journal of Pharmacological and Toxicological Methods·R RinkG N Moll
Mar 9, 2010·Peptides·Louwe de VriesMarijke Haas Jimoh Akanbi
Apr 28, 2011·Expert Opinion on Drug Metabolism & Toxicology·Auke Johan van HeelOscar P Kuipers
Oct 21, 2011·Molecular Microbiology·Rustem KhusainovOscar P Kuipers
Oct 17, 2012·Chembiochem : a European Journal of Chemical Biology·Rustem Khusainov, Oscar P Kuipers
Apr 17, 2013·Proceedings of the National Academy of Sciences of the United States of America·Neha GargWilfred A van der Donk

❮ Previous
Next ❯

Citations

Jun 16, 2015·Frontiers in Microbiology·Christophe GrangeasseIvan Mijakovic
Jan 31, 2017·Chemical Reviews·Lindsay M RepkaWilfred A van der Donk
Aug 15, 2019·Proceedings of the National Academy of Sciences of the United States of America·Ian R BothwellSatish K Nair
Dec 16, 2019·Antonie van Leeuwenhoek·Andressa FusiegerLuís Augusto Nero
Aug 17, 2021·Journal of the American Chemical Society·Alexander A VinogradovHiroaki Suga

❮ Previous
Next ❯

Datasets Mentioned

BETA
GM17

Methods Mentioned

BETA
PCR
pull-down
pull-out

Software Mentioned

HMM
UCSF chimera

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.