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Protein-glycolipid interactions studied in vitro using ESI-MS and nanodiscs: Insights into the mechanisms and energetics of binding
Download2015
Nikjah, Sanaz, Boraston, Alisdair B., Lin, Hong, Kitova, Elena N., Li, Jun, Pluvinage, Benjamin, Han, Ling Han, Klassen, John S.
Electrospray ionization-mass spectrometry (ESI-MS) analysis combined with the use of nanodiscs (NDs) to solubilize glycolipids (GLs) has recently emerged as a promising analytical method for detecting protein–GL interactions in vitro and, when applied to libraries of GLs, ranking their...
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Dissociation kinetics of the streptavidin-biotin interaction measured using direct electrospray ionization mass spectrometry analysis
Download2013
Deng, Lu, Klassen, John S., Kitova, Elena N.
Dissociation rate constants (koff) for the model high affinity interaction between biotin (B) and the homotetramer of natural core streptavidin (S4) were measured at pH 7 and temperatures ranging from 15 to 45 °C using electrospray ionization mass spectrometry (ESI-MS). Two different approaches...
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Structure and stability of carbohydrate-lipid interactions: Methylmannose polysaccharide-fatty acid complexes
Download2016
Mayer, Paul M., Richards, Michele R., Lowary, Todd L., Renaud, Justin, Klassen, John S., Tieleman, D. Peter, Kitova, Elena N., Liu, Lan, Siuda, Iwona
We report a detailed study of the structure and stability of carbohydrate–lipid interactions. Complexes of a methylmannose polysaccharide (MMP) derivative and fatty acids (FAs) served as model systems. The dependence of solution affinities and gas-phase dissociation activation energies (Ea) on FA...
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Structural basis for antibody recognition in the receptor-binding domains of toxins A and B from Clostridium difficile
Download2014
Ng, Kenneth K. S., Tanha, Jamshid, Kitova, Elena N., Eugenio, Luiz, Murase, Tomohiko, Klassen, John S., Hussack, Greg, Schorr, Melissa
Clostridium difficile infection is a serious and highly prevalent nosocomial disease in which the two large, Rho-glucosylating toxins TcdA and TcdB are the main virulence factors. We report for the first time crystal structures revealing how neutralizing and non-neutralizing single-domain...
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2015
Klassen, John S., Cairo, Christopher W.Xiong, Zi Jian, Li, Jun, Privé, Gilbert G., Leney, Aneika C., Kitova, Elena N., Darestani, Reza Rezaei, Zou, Chunxia, Nikjah, Sanaz
Protein interactions with glycolipids are implicated in diverse cellular processes. However, the study of protein–glycolipid complexes remains a significant experimental challenge. Here, we describe a powerful new assay that combines electrospray ionization mass spectrometry (ESI-MS) and...
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Quantifying protein-carbohydrate interactions using liquid sample desorption electrospray ionization mass spectrometry
Download2015
Klassen, John S., N. Kitova, Elena N., Yao, Yuyu, Shams-Ud-Doha, Km, Daneshfar, Rambod
The application of liquid sample desorption electrospray ionization mass spectrometry (liquid sample DESI-MS) for quantifying protein–carbohydrate interactions in vitro is described. Association constants for the interactions between lysozyme and β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-D-GlcNAc and...
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Identifying carbohydrate ligands of a norovirus P particle using a catch and release electrospray ionization mass spectrometry assay
Download2014
Jiang, Xi, Kitova, Elena N., Klassen, John S., Tan, Ming, Han, Ling
Noroviruses (NoVs), the major cause of epidemic acute gastroenteritis, recognize human histo-blood group antigens (HBGAs), which are present as free oligosaccharides in bodily fluid or glycolipids and glycoproteins on the surfaces of cells. The subviral P particle formed by the protruding (P)...
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2015
Tan, Ming, Jiang, Xi, Han, Ling, Kitova, Elena N., Klassen, John S., Boraston, Alisdair, B., Pluvinage, Benjamin
The binding profiles of many human noroviruses (huNoVs) for human histo-blood group antigens have been characterized. However, quantitative-binding data for these important virus–host interactions are lacking. Here, we report on the intrinsic (per binding site) affinities of HBGA oligosaccharides...
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2016
Han, Ling, Klassen, John S., Kitova, Elena N.
This study reports on the use of the catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay, combined with glycomicelles, as a method for detecting specific interactions between water-soluble proteins and glycolipids (GLs) in aqueous solution. The B subunit homopentamers...
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Carbohydrate-lipid interactions: Affinities of methylmannose polysaccharides for lipids in aqueous solution
Download2012
Xia, Li, Sun, Nian, Klassen, John S., Liu, Lan, Bai, Yu, Lowary, Todd L.
The interactions between 3-O-methyl-mannose polysaccharides (MMPs), extracted from Mycobacterium smegmatis (consisting of a mixture of MMP-10, -11, -12 and -13) or obtained by chemical synthesis (MMP-5s, -8s, -11s and -14s), and linear saturated and unsaturated fatty acids (FAs), and a commercial...