Glycoproteins and Antibodies: Solution NMR Studies

Kato, Koichi; Yamaguchi, Yoshiki

doi:10.1002/9780470034590.emrstm1233

Cited by 5 publications

(3 citation statements)

References 102 publications

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“…In addition to high glycoprotein yield, the utility of stable isotope-labeling techniques is key to the NMR analysis of glycoproteins. Several review articles [40][41][42][43] have outlined methodologies and applications of uniform and selective isotope labeling of glycoproteins, based on metabolic labeling within eukaryotic cells or enzymatic attachment of isotope-labeled monosaccharides in vitro.…”

Section: Production Of Isotopically Labeled Glycoproteins For Nmr Anamentioning

confidence: 99%

Recent advances in glycoprotein production for structural biology: toward tailored design of glycoforms

Satoh

Kato

2014

Current Opinion in Structural Biology

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Because of the complexity, heterogeneity, and flexibility of the glycans, the structural analysis of glycoproteins has been eschewed until recently, with a few prominent exceptions. This aversion may have branded structural biologists as glycophobics. However, recent technological advancements in glycoprotein expression systems, employing genetically engineered production vehicles derived from mammalian, insect, yeast, and even bacterial cells, have yielded encouraging breakthroughs. The major advance is the active control of glycoform expression of target glycoproteins based on the genetic manipulation of glycan biogenetic pathways, which was previously overlooked, abolished, or considered unmanageable. Moreover, synthetic and/or chemoenzymatic approaches now enable the preparation of glycoproteins with uniform glycoforms designed in a tailored fashion.

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Section: Production Of Isotopically Labeled Glycoproteins For Nmr Anamentioning

confidence: 99%

Recent advances in glycoprotein production for structural biology: toward tailored design of glycoforms

Satoh

Kato

2014

Current Opinion in Structural Biology

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“…Thus, detailed characterization of serum protein interactions is desirable for the development of biopharmaceuticals with minimal side effects. However, detailed structural characterizations of antibody interactions have thus been performed almost exclusively under homogeneous in vitro conditions, employing crystallographic and spectroscopic techniques [ 8 , 9 , 10 ].…”

Section: Introductionmentioning

confidence: 99%

NMR Detection of Semi-Specific Antibody Interactions in Serum Environments

Yanaka

Yamazaki²,

Yogo

et al. 2017

Molecules

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Although antibody functions are executed in heterogeneous blood streams characterized by molecular crowding and promiscuous intermolecular interaction, detailed structural characterizations of antibody interactions have thus far been performed under homogeneous in vitro conditions. NMR spectroscopy potentially has the ability to study protein structures in heterogeneous environments, assuming that the target protein can be labeled with NMR-active isotopes. Based on our successful development of isotope labeling of antibody glycoproteins, here we apply NMR spectroscopy to characterize antibody interactions in heterogeneous extracellular environments using mouse IgG-Fc as a test molecule. In human serum, many of the HSQC peaks originating from the Fc backbone exhibited attenuation in intensity of various magnitudes. Similar spectral changes were induced by the Fab fragment of polyclonal IgG isolated from the serum, but not by serum albumin, indicating that a subset of antibodies reactive with mouse IgG-Fc exists in human serum without preimmunization. The metaepitopes recognized by serum polyclonal IgG cover the entire molecular surface of Fc, including the binding sites to Fc receptors and C1q. In-serum NMR observation will offer useful tools for the detailed characterization of biopharamaceuticals, including therapeutic antibodies in physiologically relevant heterogeneous environments, also giving deeper insight into molecular recognition by polyclonal antibodies in the immune system.

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“…NMR spectroscopy is a useful tool for the conformational analysis of oligosaccharides in solution [ 6 , 7 , 8 , 9 ]. However, it is not feasible to determine the dynamic conformation of oligosaccharides solely on the basis of local conformational restraints derived from nuclear Overhauser effects (NOEs) and scalar couplings.…”

Section: Introductionmentioning

confidence: 99%

Application of Paramagnetic NMR-Validated Molecular Dynamics Simulation to the Analysis of a Conformational Ensemble of a Branched Oligosaccharide

et al. 2012

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Oligosaccharides of biological importance often exhibit branched covalent structures and dynamic conformational multiplicities. Here we report the application of a method that we developed, which combined molecular dynamics (MD) simulations and lanthanide-assisted paramagnetic NMR spectroscopy, to evaluate the dynamic conformational ensemble of a branched oligosaccharide. A lanthanide-chelating tag was attached to the reducing end of the branched tetrasaccharide of GM2 ganglioside to observe pseudocontact shifts as the source of long distance information for validating the conformational ensemble derived from MD simulations. By inspecting the results, the conformational space of the GM2 tetrasaccharide was compared with that of its nonbranched derivative, the GM3 trisaccharide.

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Glycoproteins and Antibodies: Solution NMR Studies

Cited by 5 publications

References 102 publications

Recent advances in glycoprotein production for structural biology: toward tailored design of glycoforms

Recent advances in glycoprotein production for structural biology: toward tailored design of glycoforms

NMR Detection of Semi-Specific Antibody Interactions in Serum Environments

Application of Paramagnetic NMR-Validated Molecular Dynamics Simulation to the Analysis of a Conformational Ensemble of a Branched Oligosaccharide

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