Oriented immobilization of peptide-N-glycosidase F on a
monolithic support for glycosylation analysis

J 2013

Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis

KRENKOVA, Jana, Akos SZEKRENYES, Zsolt KERESZTESSY, František FORET, Andras GUTTMAN et. al.

Základní údaje

Originální název

Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis

Autoři

KRENKOVA, Jana (203 Česká republika), Akos SZEKRENYES (348 Maďarsko), Zsolt KERESZTESSY (348 Maďarsko), František FORET (203 Česká republika, garant, domácí) a Andras GUTTMAN (348 Maďarsko)

Vydání

Journal of Chromatography A, AMSTERDAM, Elsevier, 2013, 0021-9673

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 4.258

Kód RIV

RIV/00216224:14740/13:00072093

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1016/j.chroma.2013.10.087

UT WoS

000328717200007

Klíčová slova anglicky

Enzyme microreactor; Oriented immobilization; Monolith; PNGase F; Deglycosylation

Štítky

ok, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 14. 2. 2014 00:35, Olga Křížová

Anotace

V originále

In this paper, we report on a novel oriented peptide-N-glycosidase F (PNGase F) immobilization approach onto methacrylate based monolithic support for rapid, reproducible and efficient release of the N-linked carbohydrate moieties from glycoproteins. The glutathione-S-transferase-fusion PNGase F (PNGase F-GST) was expressed in Escherichia coli using regular vector technology. The monolithic pore surface was functionalized with glutathione via a succinimidy1-6-(iodoacetyl-amino)-hexanoate linker and the specific affinity of GST toward glutathione was utilized for the oriented coupling. This novel immobilization procedure was compared with reductive amination technique commonly used for non-oriented enzyme immobilization via primary amine functionalities. Both coupling approaches were compared using enzymatic treatment of several glycoproteins, such as ribonuclease B, fetuin and immunoglobulin G followed by MALDI/MS and CE-LIF analysis of the released glycans. Orientedly immobilized PNGase F via GST-glutathione coupling showed significantly higher activity, remained stable for several months, and allowed rapid release of various types of glycans (high-mannose, core fucosylated, sialylated, etc.) from glycoproteins. Complete protein deglycosylation was obtained as fast as in several seconds when using flow-through immobilized microreactors. (C) 2013 Elsevier B.V All rights reserved.

Návaznosti

ED1.1.00/02.0068, projekt VaV

Název: CEITEC - central european institute of technology

Citovat

KRENKOVA, Jana, Akos SZEKRENYES, Zsolt KERESZTESSY, František FORET a Andras GUTTMAN. Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis. Journal of Chromatography A. AMSTERDAM: Elsevier, 2013, roč. 1322, Dec, s. 54-61. ISSN 0021-9673. Dostupné z: https://dx.doi.org/10.1016/j.chroma.2013.10.087.

@article{1165396,
   author = {Krenkova, Jana and Szekrenyes, Akos and Keresztessy, Zsolt and Foret, František and Guttman, Andras},
   article_location = {AMSTERDAM},
   article_number = {Dec},
   doi = {http://dx.doi.org/10.1016/j.chroma.2013.10.087},
   keywords = {Enzyme microreactor; Oriented immobilization; Monolith; PNGase F; Deglycosylation},
   language = {eng},
   issn = {0021-9673},
   journal = {Journal of Chromatography A},
   title = {Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis},
   volume = {1322},
   year = {2013}
}

TY  - JOUR
ID  - 1165396
AU  - Krenkova, Jana - Szekrenyes, Akos - Keresztessy, Zsolt - Foret, František - Guttman, Andras
PY  - 2013
TI  - Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis
JF  - Journal of Chromatography A
VL  - 1322
IS  - Dec
SP  - 54-61
EP  - 54-61
PB  - Elsevier
SN  - 00219673
KW  - Enzyme microreactor
KW  - Oriented immobilization
KW  - Monolith
KW  - PNGase F
KW  - Deglycosylation
N2  - In this paper, we report on a novel oriented peptide-N-glycosidase F (PNGase F) immobilization approach onto methacrylate based monolithic support for rapid, reproducible and efficient release of the N-linked carbohydrate moieties from glycoproteins. The glutathione-S-transferase-fusion PNGase F (PNGase F-GST) was expressed in Escherichia coli using regular vector technology. The monolithic pore surface was functionalized with glutathione via a succinimidy1-6-(iodoacetyl-amino)-hexanoate linker and the specific affinity of GST toward glutathione was utilized for the oriented coupling. This novel immobilization procedure was compared with reductive amination technique commonly used for non-oriented enzyme immobilization via primary amine functionalities. Both coupling approaches were compared using enzymatic treatment of several glycoproteins, such as ribonuclease B, fetuin and immunoglobulin G followed by MALDI/MS and CE-LIF analysis of the released glycans. Orientedly immobilized PNGase F via GST-glutathione coupling showed significantly higher activity, remained stable for several months, and allowed rapid release of various types of glycans (high-mannose, core fucosylated, sialylated, etc.) from glycoproteins. Complete protein deglycosylation was obtained as fast as in several seconds when using flow-through immobilized microreactors. (C) 2013 Elsevier B.V All rights reserved.
ER  -

KRENKOVA, Jana, Akos SZEKRENYES, Zsolt KERESZTESSY, František FORET a Andras GUTTMAN. Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis. \textit{Journal of Chromatography A}. AMSTERDAM: Elsevier, 2013, roč.~1322, Dec, s.~54-61. ISSN~0021-9673. Dostupné z: https://dx.doi.org/10.1016/j.chroma.2013.10.087.

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