Biochemical characterization of WbdN, a β1,3-glucosyltransferase involved in O-antigen synthesis in enterohemorrhagic Escherichia coli O157

Glycobiology
Yin GaoInka Brockhausen

Abstract

The enterohemorrhagic O157 strain of Escherichia coli, which is one of the most well-known bacterial pathogens, has an O-antigen repeating unit structure with the sequence [-2-d-Rha4NAcα1-3-l-Fucα1-4-d-Glcβ1-3-d-GalNAcα1-]. The O-antigen gene cluster of E. coli O157 contains the genes responsible for the assembly of this repeating unit and includes wbdN. In spite of cloning many O-antigen genes, biochemical characterization has been done on very few enzymes involved in O-antigen synthesis. In this work, we expressed the wbdN gene in E. coli BL21, and the His-tagged protein was purified. WbdN activity was characterized using the donor substrate UDP-[(14)C]Glc and the synthetic acceptor substrate GalNAcα-O-PO(3)-PO(3)-(CH(2))(11)-O-Ph. The enzyme product was isolated by high pressure liquid chromatography, and mass spectrometry showed that one Glc residue was transferred to the acceptor by WbdN. Nuclear magnetic resonance analysis of the product structure indicated that Glc was β1-3 linked to GalNAc. WbdN contains a conserved DxD motif and requires divalent metal ions for full activity. WbdN activity has an optimal pH between 7 and 8 and is highly specific for UDP-Glc as the donor substrate. GalNAcα derivatives lacking the diphos...Continue Reading

References

Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Sep 1, 1990·Journal of Bacteriology·R C Goldman, F Hunt
Jun 1, 1986·Journal of Bacteriology·P J HitchcockD C Morrison
May 1, 1995·Trends in Microbiology·C Whitfield
Dec 1, 1996·Trends in Microbiology·P R ReevesP D Rick
May 1, 1997·Infection and Immunity·E A Dean-NystromH W Moon
Feb 11, 1998·Clinical Microbiology Reviews·J P Nataro, J B Kaper
Oct 24, 2000·Current Opinion in Structural Biology·U M Unligil, J M Rini
Jun 5, 2002·Annual Review of Biochemistry·Christian R H Raetz, Chris Whitfield
Apr 18, 2003·Frontiers in Bioscience : a Journal and Virtual Library·Miguel A Valvano
Jun 19, 2003·Journal of Bacteriology·Els van den Brink-van der LaanBen de Kruijff
Dec 13, 2003·Carbohydrate Research·Martine Caroff, Doris Karibian
Feb 3, 2005·Bioorganic & Medicinal Chemistry Letters·Pedro J Montoya-PeleazInka Brockhausen
Jul 23, 2005·Glycobiology·Christelle BretonAnne Imberty
Apr 1, 2006·Seminars in Thrombosis and Hemostasis·Peter F ZipfelChristine Skerka
May 30, 2006·Current Microbiology·Narelle FeganKari Steven Gobius
Aug 21, 2007·Clinical Journal of the American Society of Nephrology : CJASN·Elena BresinUNKNOWN International Registry of Recurrent and Familial HUS/TTP
Nov 29, 2007·Bioorganic & Medicinal Chemistry Letters·Inka BrockhausenOle Hindsgaul
Apr 30, 2009·Journal of Molecular Recognition : JMR·Joshua StraussTerri A Camesano
Nov 20, 2009·The Journal of Biological Chemistry·Jeffrey S RushCharles J Waechter
Dec 2, 2009·Applied and Environmental Microbiology·Lydia V RumpSteven R Monday
Jan 25, 2011·Proceedings of the National Academy of Sciences of the United States of America·Elizaveta FreinkmanDaniel Kahne

❮ Previous
Next ❯

Citations

Oct 13, 2012·Journal of the American Chemical Society·Moritz Winger, Mark von Itzstein
Nov 5, 2014·Frontiers in Immunology·Inka Brockhausen
Nov 30, 2019·FEMS Microbiology Reviews·Bin LiuGöran Widmalm

❮ 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.