Abstract:
:Glycoproteins are characterized by the heterogeneous and dynamic nature of their glycan moieties, which hamper crystallographic analysis. NMR spectroscopy provides potential advantages in dealing with such complicated systems, given that the target molecules can be isotopically labeled. Methods of metabolic isotope labeling in recombinant glycoproteins have been developed recently using a variety of eukaryotic production vehicles, including mammalian, yeast, insect, and plant cells, each of which has a distinct N-glycan diversification pathway. Yeast genetic engineering has enabled the overexpression of homogeneous high-mannose-type oligosaccharides with 13C labeling for NMR characterization of their conformational dynamics. The utility of stable isotope-assisted NMR spectroscopy has also been demonstrated using the Fc fragment of immunoglobulin G (IgG) as a model glycoprotein, providing useful information regarding intramolecular carbohydrate-protein interactions. Transverse relaxation optimization of intact IgG with a molecular mass of 150 kDa has been achieved by tailored deuteration of selected amino acid residues using a mammalian expression system. This offers a useful probe for the characterization of molecular interaction networks in multimolecular crowded systems typified by serum. Perspectives regarding the development of techniques for tailoring glycoform designs and isotope labeling of recombinant glycoproteins are also discussed.
journal_name
J Biomol NMRjournal_title
Journal of biomolecular NMRauthors
Yanaka S,Yagi H,Yogo R,Yagi-Utsumi M,Kato Kdoi
10.1007/s10858-018-0169-2subject
Has Abstractpub_date
2018-07-01 00:00:00pages
193-202issue
3eissn
0925-2738issn
1573-5001pii
10.1007/s10858-018-0169-2journal_volume
71pub_type
杂志文章abstract::NifU-like proteins are a highly conserved protein that serves as the scaffold for assembly of Fe-S clusters. Chloroplastic NifU-like proteins have tandem NifU like domains, named domain I and domain II. Although the amino acid sequences of these domains are very similar to each other, the predicted functional region f...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-007-9155-9
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-011-9515-3
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2019-02-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-019-00266-0
更新日期:2019-11-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-009-9335-x
更新日期:2009-08-01 00:00:00
abstract::Sample solubility is essential for structural studies of proteins by solution NMR. Attachment of a solubility enhancement tag, such as GB1, MBP and thioredoxin, to a target protein has been used for this purpose. However, signal overlap of the tag with the target protein often made the spectral analysis difficult. Her...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-008-9296-5
更新日期:2009-03-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-015-9976-x
更新日期:2015-10-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1023/a:1023833407515
更新日期:2003-07-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1023/a:1008340210079
更新日期:2001-02-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1023/b:jnmr.0000048948.66235.d9
更新日期:2004-10-01 00:00:00
abstract::Carr-Purcell-Meiboom-Gill relaxation dispersion (CPMG RD) NMR spectroscopy has emerged as a powerful tool for quantifying the kinetics and thermodynamics of millisecond time-scale exchange processes involving the interconversion between a visible ground state and one or more minor, sparsely populated invisible 'excite...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-012-9617-6
更新日期:2012-05-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00156619
更新日期:1994-07-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF02192865
更新日期:1991-11-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1023/a:1008308128050
更新日期:1999-06-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-015-9938-3
更新日期:2015-06-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-019-00274-0
更新日期:2019-11-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-013-9776-0
更新日期:2013-10-01 00:00:00
abstract::The efficiency of cell-free protein synthesis combined with combinatorial selective 15N-labelling provides a method for the rapid assignment of 15N-HSQC cross-peaks to the 19 different non-proline amino-acid types from five 15N-HSQC spectra. This strategy was explored with two different constructs of the C-terminal do...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-005-5021-9
更新日期:2006-01-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2016-08-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/BF00197809
更新日期:1995-11-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
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更新日期:2012-02-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-018-0187-0
更新日期:2018-05-01 00:00:00
abstract::We perform a detailed comparison of fast backbone dynamics probed at amide nitrogen versus carbonyl carbon sites for dematin headpiece C-terminal domain (DHP) and its S74E mutant (DHPS74E). Carbonyl dynamics is probed via auto-correlated longitudinal rates and transverse C'/C'-C(alpha) CSA/dipolar and C'/C'-N CSA/dipo...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-010-9417-9
更新日期:2010-06-01 00:00:00
abstract::We demonstrate a novel sparse (13)C labelling approach for methylotrophic yeast P. pastoris expression system, towards solid-state NMR studies of eukaryotic membrane proteins. The labelling scheme was achieved by co-utilizing natural abundance methanol and specifically (13)C labelled glycerol as carbon sources in the ...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-016-0033-1
更新日期:2016-05-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-005-3592-0
更新日期:2005-05-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-005-1846-5
更新日期:2004-11-01 00:00:00
abstract::Artifacts associated with the measurement of methyl (1)H single quantum CPMG-based relaxation dispersion profiles are described. These artifacts arise due to the combination of cross-correlated spin relaxation effects involving intra-methyl (1)H-(1)H dipolar interactions and imperfections in (1)H refocusing pulses tha...
journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-005-2468-7
更新日期:2005-04-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-018-0207-0
更新日期:2018-10-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-014-9856-9
更新日期:2014-11-01 00:00:00
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journal_title:Journal of biomolecular NMR
pub_type: 杂志文章
doi:10.1007/s10858-011-9583-4
更新日期:2012-01-01 00:00:00