The wbnH gene of Escherichia coli O86:H2 encodes an alpha-1,3-N-acetylgalactosaminyl transferase involved in the O-repeating unit biosynthesis

Biochemical and Biophysical Research Communications
Wen YiPeng George Wang

Abstract

O-repeating unit biosynthesis is the first committed step in lipopolysaccharide (LPS) biosynthesis in a variety of gram-negative bacteria. The wbnH gene was previously proposed to encode a glycosyltransferase involved in O-repeating unit synthesis in Escherichia coli O86:H2 strain. In this work, we provide biochemical evidence to show that wbnH encodes a N-acetylgalactosaminyl transferase (GalNAcT) that catalyzes the transfer of GalNAc from UDP-GalNAc to the GalNAc-pyrophosphate-lipid acceptor. WbnH activity was characterized using a synthetic acceptor substrate GalNAc alpha-PP-O(CH2)11-OPh. The resulting disaccharide product GalNAc-alpha-1,3-GalNAc alpha-PP-O(CH2)11-OPh was analyzed by LC-MS and NMR spectroscopy. Substrate specificity study indicates that pyrophosphate and hydrophobic lipid moiety are structural requirements for WbnH activity. Divalent metal cations are not required for enzyme catalysis, suggesting WbnH belongs to glycosyltransferase GT-B superfamily. Our results complete the characterization of O86 O-unit assembly pathway, and provide the access of chemically defined O-unit substrates for the further investigation of O-antigen biosynthetic mechanism.

References

May 1, 1985·Journal of Bacteriology·P D RickD Bitter-Suermann
Apr 1, 1997·FEMS Microbiology Letters·E García, R López
May 17, 2000·Proceedings of the National Academy of Sciences of the United States of America·D YuD L Court
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Aug 29, 2000·Research in Microbiology·E GarcíaR López
May 23, 2002·Biochemistry·Lan ChenSuzanne Walker
Jun 5, 2002·Annual Review of Biochemistry·Christian R H Raetz, Chris Whitfield
Dec 10, 2002·Current Opinion in Chemical Biology·Kristi Lazar, Suzanne Walker
Dec 25, 2002·Chemistry & Biology·Yanan Hu, Suzanne Walker
Jan 23, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yanan HuSuzanne Walker
Mar 31, 2004·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·M Stephen Trent
Feb 3, 2005·Bioorganic & Medicinal Chemistry Letters·Pedro J Montoya-PeleazInka Brockhausen
Dec 14, 2006·ACS Chemical Biology·Cynthia GinsbergSuzanne Walker
Dec 20, 2008·Journal of Lipid Research·Teresa A GarrettRebecca L Rose

❮ Previous
Next ❯

Citations

Apr 27, 2010·Nature Chemical Biology·Robert WoodwardPeng George Wang
Apr 1, 2015·Protein & Cell·Fengzhi LiYuequan Shen
Jul 14, 2011·Chemical Reviews·Ryan M SchmaltzChi-Huey Wong
May 29, 2014·Journal of the American Chemical Society·Mario A Martinez FariasLaura L Kiessling
Nov 29, 2007·Bioorganic & Medicinal Chemistry Letters·Inka BrockhausenOle Hindsgaul

❮ 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.