Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells

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Standard

Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells. / Schjoldager, Katrine Ter-Borch Gram; Vakhrushev, Sergey Y; Kong, Yun; Steentoft, Catharina; Nudelman, Aaron S; Pedersen, Nis Borbye; Wandall, Hans H; Mandel, Ulla; Bennett, Eric P; Levery, Steven B; Clausen, Henrik.

I: Proceedings of the National Academy of Sciences USA (PNAS), Bind 109, Nr. 25, 06.2012, s. 9893-8.

Publikation: Bidrag til tidsskriftTidsskriftartikelfagfællebedømt

Harvard

Schjoldager, KT-BG, Vakhrushev, SY, Kong, Y, Steentoft, C, Nudelman, AS, Pedersen, NB, Wandall, HH, Mandel, U, Bennett, EP, Levery, SB & Clausen, H 2012, 'Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells', Proceedings of the National Academy of Sciences USA (PNAS), bind 109, nr. 25, s. 9893-8. https://doi.org/10.1073/pnas.1203563109

APA

Schjoldager, K. T-B. G., Vakhrushev, S. Y., Kong, Y., Steentoft, C., Nudelman, A. S., Pedersen, N. B., Wandall, H. H., Mandel, U., Bennett, E. P., Levery, S. B., & Clausen, H. (2012). Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells. Proceedings of the National Academy of Sciences USA (PNAS), 109(25), 9893-8. https://doi.org/10.1073/pnas.1203563109

Vancouver

Schjoldager KT-BG, Vakhrushev SY, Kong Y, Steentoft C, Nudelman AS, Pedersen NB o.a. Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells. Proceedings of the National Academy of Sciences USA (PNAS). 2012 jun.;109(25):9893-8. https://doi.org/10.1073/pnas.1203563109

Author

Schjoldager, Katrine Ter-Borch Gram ; Vakhrushev, Sergey Y ; Kong, Yun ; Steentoft, Catharina ; Nudelman, Aaron S ; Pedersen, Nis Borbye ; Wandall, Hans H ; Mandel, Ulla ; Bennett, Eric P ; Levery, Steven B ; Clausen, Henrik. / Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells. I: Proceedings of the National Academy of Sciences USA (PNAS). 2012 ; Bind 109, Nr. 25. s. 9893-8.

Bibtex

@article{4a61d735584f4e8f9904e3e04ab9b549,
title = "Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells",
abstract = "Our knowledge of the O-glycoproteome [N-acetylgalactosamine (GalNAc) type] is highly limited. The O-glycoproteome is differentially regulated in cells by dynamic expression of a subset of 20 polypeptide GalNAc-transferases (GalNAc-Ts), and methods to identify important functions of individual GalNAc-Ts are largely unavailable. We recently introduced SimpleCells, i.e., human cell lines made deficient in O-glycan extension by zinc finger nuclease targeting of a key gene in O-glycan elongation (Cosmc), which allows for proteome-wide discovery of O-glycoproteins. Here we have extended the SimpleCell concept to include proteome-wide discovery of unique functions of individual GalNAc-Ts. We used the GalNAc-T2 isoform implicated in dyslipidemia and the human HepG2 liver cell line to demonstrate unique functions of this isoform. We confirm that GalNAc-T2-directed site-specific O-glycosylation inhibits proprotein activation of the lipase inhibitor ANGPTL3 in HepG2 cells and further identify eight O-glycoproteins exclusively glycosylated by T2 of which one, ApoC-III, is implicated in dyslipidemia. Our study supports an essential role for GalNAc-T2 in lipid metabolism, provides serum biomarkers for GalNAc-T2 enzyme function, and validates the use of GALNT gene targeting with SimpleCells for broad discovery of disease-causing deficiencies in O-glycosylation. The presented glycoengineering strategy opens the way for proteome-wide discovery of functions of GalNAc-T isoforms and their role in congenital diseases and disorders.",
author = "Schjoldager, {Katrine Ter-Borch Gram} and Vakhrushev, {Sergey Y} and Yun Kong and Catharina Steentoft and Nudelman, {Aaron S} and Pedersen, {Nis Borbye} and Wandall, {Hans H} and Ulla Mandel and Bennett, {Eric P} and Levery, {Steven B} and Henrik Clausen",
year = "2012",
month = jun,
doi = "10.1073/pnas.1203563109",
language = "English",
volume = "109",
pages = "9893--8",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "The National Academy of Sciences of the United States of America",
number = "25",

}

RIS

TY - JOUR

T1 - Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells

AU - Schjoldager, Katrine Ter-Borch Gram

AU - Vakhrushev, Sergey Y

AU - Kong, Yun

AU - Steentoft, Catharina

AU - Nudelman, Aaron S

AU - Pedersen, Nis Borbye

AU - Wandall, Hans H

AU - Mandel, Ulla

AU - Bennett, Eric P

AU - Levery, Steven B

AU - Clausen, Henrik

PY - 2012/6

Y1 - 2012/6

N2 - Our knowledge of the O-glycoproteome [N-acetylgalactosamine (GalNAc) type] is highly limited. The O-glycoproteome is differentially regulated in cells by dynamic expression of a subset of 20 polypeptide GalNAc-transferases (GalNAc-Ts), and methods to identify important functions of individual GalNAc-Ts are largely unavailable. We recently introduced SimpleCells, i.e., human cell lines made deficient in O-glycan extension by zinc finger nuclease targeting of a key gene in O-glycan elongation (Cosmc), which allows for proteome-wide discovery of O-glycoproteins. Here we have extended the SimpleCell concept to include proteome-wide discovery of unique functions of individual GalNAc-Ts. We used the GalNAc-T2 isoform implicated in dyslipidemia and the human HepG2 liver cell line to demonstrate unique functions of this isoform. We confirm that GalNAc-T2-directed site-specific O-glycosylation inhibits proprotein activation of the lipase inhibitor ANGPTL3 in HepG2 cells and further identify eight O-glycoproteins exclusively glycosylated by T2 of which one, ApoC-III, is implicated in dyslipidemia. Our study supports an essential role for GalNAc-T2 in lipid metabolism, provides serum biomarkers for GalNAc-T2 enzyme function, and validates the use of GALNT gene targeting with SimpleCells for broad discovery of disease-causing deficiencies in O-glycosylation. The presented glycoengineering strategy opens the way for proteome-wide discovery of functions of GalNAc-T isoforms and their role in congenital diseases and disorders.

AB - Our knowledge of the O-glycoproteome [N-acetylgalactosamine (GalNAc) type] is highly limited. The O-glycoproteome is differentially regulated in cells by dynamic expression of a subset of 20 polypeptide GalNAc-transferases (GalNAc-Ts), and methods to identify important functions of individual GalNAc-Ts are largely unavailable. We recently introduced SimpleCells, i.e., human cell lines made deficient in O-glycan extension by zinc finger nuclease targeting of a key gene in O-glycan elongation (Cosmc), which allows for proteome-wide discovery of O-glycoproteins. Here we have extended the SimpleCell concept to include proteome-wide discovery of unique functions of individual GalNAc-Ts. We used the GalNAc-T2 isoform implicated in dyslipidemia and the human HepG2 liver cell line to demonstrate unique functions of this isoform. We confirm that GalNAc-T2-directed site-specific O-glycosylation inhibits proprotein activation of the lipase inhibitor ANGPTL3 in HepG2 cells and further identify eight O-glycoproteins exclusively glycosylated by T2 of which one, ApoC-III, is implicated in dyslipidemia. Our study supports an essential role for GalNAc-T2 in lipid metabolism, provides serum biomarkers for GalNAc-T2 enzyme function, and validates the use of GALNT gene targeting with SimpleCells for broad discovery of disease-causing deficiencies in O-glycosylation. The presented glycoengineering strategy opens the way for proteome-wide discovery of functions of GalNAc-T isoforms and their role in congenital diseases and disorders.

U2 - 10.1073/pnas.1203563109

DO - 10.1073/pnas.1203563109

M3 - Journal article

C2 - 22566642

VL - 109

SP - 9893

EP - 9898

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 25

ER -

ID: 38331819