Efficient Procedure for N-Glycan Analyses and Detection of Endo
H-Like Activity in Human Tumor Specimens

J 2016

Efficient Procedure for N-Glycan Analyses and Detection of Endo H-Like Activity in Human Tumor Specimens

LATTOVÁ, Erika, Joseph BRYANT, Jana SKŘIČKOVÁ, Zbyněk ZDRÁHAL, Mikuláš POPOVIČ et. al.

Základní údaje

Originální název

Efficient Procedure for N-Glycan Analyses and Detection of Endo H-Like Activity in Human Tumor Specimens

Autoři

LATTOVÁ, Erika (703 Slovensko, domácí), Joseph BRYANT (840 Spojené státy), Jana SKŘIČKOVÁ (203 Česká republika, domácí), Zbyněk ZDRÁHAL (203 Česká republika, garant, domácí) a Mikuláš POPOVIČ (703 Slovensko)

Vydání

JOURNAL OF PROTEOME RESEARCH, Washington, AMER CHEMICAL SOC, 2016, 1535-3893

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.268

Kód RIV

RIV/00216224:14740/16:00088140

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1021/acs.jproteome.6b00346

UT WoS

000381235900039

Klíčová slova anglicky

Biopsy; cancer cells; EndoH; glycans; glycosylation; mass spectrometry; tumor tissue

Štítky

rivok

Změněno: 21. 2. 2017 15:37, Mgr. Eva Špillingová

Anotace

V originále

Although the importance of glycosylation has been thoroughly recognized in association with a number of biological processes, efficient assessments of glycans have been hampered by both the limited size of specimens and lengthy sample preparations, particularly in clinical settings. Here we report a simple preparative method for N-glycan analyses. It involves only short one-step chloroform methanol extraction in presence or absence of water prior to PNGase F deglycosylation. The procedure was successfully applied to the investigation of N-glycans obtained from small numbers of in vitro cultured cancer cells (<= 1 x 10(5)) and to tumor tissues, including patient biopsies of small size. MALDI-MS analysis confirmed the efficient release of all N-glycan types including complex forms with poly-N-acetyllactosamine chains. In addition, nonaqueous extraction of specimens from several established cancer cell lines, as well as patient tumor tissues, yielded high-mannose glycans with one G1cNAc moiety (Man(3-9)GlcNAc), strongly suggesting preservation of enzymatic activity analogous to Endo H enzyme. In summary, the method is both a step toward the practical use of glycan profiling and a way to detect Endo H-like activity in cancer specimens.

Návaznosti

GBP206/12/G151, projekt VaV

Název: Centrum nových přístupů k bioanalýze a molekulární diagnostice

LM2015043, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii (Akronym: CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech Infrastructure for Integrative Structural Biology

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CEITEC 2020

Citovat

LATTOVÁ, Erika, Joseph BRYANT, Jana SKŘIČKOVÁ, Zbyněk ZDRÁHAL a Mikuláš POPOVIČ. Efficient Procedure for N-Glycan Analyses and Detection of Endo H-Like Activity in Human Tumor Specimens. JOURNAL OF PROTEOME RESEARCH. Washington: AMER CHEMICAL SOC, 2016, roč. 15, č. 8, s. 2777-2786. ISSN 1535-3893. Dostupné z: https://dx.doi.org/10.1021/acs.jproteome.6b00346.

@article{1353957,
   author = {Lattová, Erika and Bryant, Joseph and Skřičková, Jana and Zdráhal, Zbyněk and Popovič, Mikuláš},
   article_location = {Washington},
   article_number = {8},
   doi = {http://dx.doi.org/10.1021/acs.jproteome.6b00346},
   keywords = {Biopsy; cancer cells; EndoH; glycans; glycosylation; mass spectrometry; tumor tissue},
   language = {eng},
   issn = {1535-3893},
   journal = {JOURNAL OF PROTEOME RESEARCH},
   title = {Efficient Procedure for N-Glycan Analyses and Detection of Endo H-Like Activity in Human Tumor Specimens},
   url = {http://pubs.acs.org/doi/abs/10.1021/acs.jproteome.6b00346},
   volume = {15},
   year = {2016}
}

TY  - JOUR
ID  - 1353957
AU  - Lattová, Erika - Bryant, Joseph - Skřičková, Jana - Zdráhal, Zbyněk - Popovič, Mikuláš
PY  - 2016
TI  - Efficient Procedure for N-Glycan Analyses and Detection of Endo H-Like Activity in Human Tumor Specimens
JF  - JOURNAL OF PROTEOME RESEARCH
VL  - 15
IS  - 8
SP  - 2777-2786
EP  - 2777-2786
PB  - AMER CHEMICAL SOC
SN  - 15353893
KW  - Biopsy
KW  - cancer cells
KW  - EndoH
KW  - glycans
KW  - glycosylation
KW  - mass spectrometry
KW  - tumor tissue
UR  - http://pubs.acs.org/doi/abs/10.1021/acs.jproteome.6b00346
L2  - http://pubs.acs.org/doi/abs/10.1021/acs.jproteome.6b00346
N2  - Although the importance of glycosylation has been thoroughly recognized in association with a number of biological processes, efficient assessments of glycans have been hampered by both the limited size of specimens and lengthy sample preparations, particularly in clinical settings. Here we report a simple preparative method for N-glycan analyses. It involves only short one-step chloroform methanol extraction in presence or absence of water prior to PNGase F deglycosylation. The procedure was successfully applied to the investigation of N-glycans obtained from small numbers of in vitro cultured cancer cells (<= 1 x 10(5)) and to tumor tissues, including patient biopsies of small size. MALDI-MS analysis confirmed the efficient release of all N-glycan types including complex forms with poly-N-acetyllactosamine chains. In addition, nonaqueous extraction of specimens from several established cancer cell lines, as well as patient tumor tissues, yielded high-mannose glycans with one G1cNAc moiety (Man(3-9)GlcNAc), strongly suggesting preservation of enzymatic activity analogous to Endo H enzyme. In summary, the method is both a step toward the practical use of glycan profiling and a way to detect Endo H-like activity in cancer specimens.
ER  -

LATTOVÁ, Erika, Joseph BRYANT, Jana SKŘIČKOVÁ, Zbyněk ZDRÁHAL a Mikuláš POPOVIČ. Efficient Procedure for N-Glycan Analyses and Detection of Endo H-Like Activity in Human Tumor Specimens. \textit{JOURNAL OF PROTEOME RESEARCH}. Washington: AMER CHEMICAL SOC, 2016, roč.~15, č.~8, s.~2777-2786. ISSN~1535-3893. Dostupné z: https://dx.doi.org/10.1021/acs.jproteome.6b00346.

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