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.

Basic information

Original name

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

Authors

LATTOVÁ, Erika (703 Slovakia, belonging to the institution), Joseph BRYANT (840 United States of America), Jana SKŘIČKOVÁ (203 Czech Republic, belonging to the institution), Zbyněk ZDRÁHAL (203 Czech Republic, guarantor, belonging to the institution) and Mikuláš POPOVIČ (703 Slovakia)

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 4.268

RIV identification code

RIV/00216224:14740/16:00088140

Organization unit

Central European Institute of Technology

DOI

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

UT WoS

000381235900039

Keywords in English

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

Abstract

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.

Links

GBP206/12/G151, research and development project

Name: Centrum nových přístupů k bioanalýze a molekulární diagnostice

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

LATTOVÁ, Erika, Joseph BRYANT, Jana SKŘIČKOVÁ, Zbyněk ZDRÁHAL and 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, vol. 15, No 8, p. 2777-2786. ISSN 1535-3893. Available from: 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 and 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, vol.~15, No~8, p.~2777-2786. ISSN~1535-3893. Available from: https://dx.doi.org/10.1021/acs.jproteome.6b00346.

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