Analysis of Site-specific N-glycan Remodeling in the Endoplasmic Reticulum and the Golgi

Overview

Journal Glycobiology

Date 2015 Aug 5

PMID 26240167

Citations 28

Authors

Ivan Hang

Chia-Wei Lin

Oliver C Grant

Susanna Fleurkens

Thomas K Villiger

Miroslav Soos

Massimo Morbidelli

Robert J Woods

Robert Gauss

Markus Aebi

Affiliations

Soon will be listed here.

Abstract

The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing.

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References

Parekh R, Tse A, Dwek R, Williams A, Rademacher T . Tissue-specific N-glycosylation, site-specific oligosaccharide patterns and lentil lectin recognition of rat Thy-1. EMBO J. 1987; 6(5):1233-44. PMC: 553924. DOI: 10.1002/j.1460-2075.1987.tb02359.x. View

Krokhin O, Ens W, Standing K, Wilkins J, Perreault H . Site-specific N-glycosylation analysis: matrix-assisted laser desorption/ionization quadrupole-quadrupole time-of-flight tandem mass spectral signatures for recognition and identification of glycopeptides. Rapid Commun Mass Spectrom. 2004; 18(18):2020-30. PMC: 7169070. DOI: 10.1002/rcm.1585. View

Ferrara C, Grau S, Jager C, Sondermann P, Brunker P, Waldhauer I . Unique carbohydrate-carbohydrate interactions are required for high affinity binding between FcgammaRIII and antibodies lacking core fucose. Proc Natl Acad Sci U S A. 2011; 108(31):12669-74. PMC: 3150898. DOI: 10.1073/pnas.1108455108. View

Niwa R, Shoji-Hosaka E, Sakurada M, Shinkawa T, Uchida K, Nakamura K . Defucosylated chimeric anti-CC chemokine receptor 4 IgG1 with enhanced antibody-dependent cellular cytotoxicity shows potent therapeutic activity to T-cell leukemia and lymphoma. Cancer Res. 2004; 64(6):2127-33. DOI: 10.1158/0008-5472.can-03-2068. View

Sazinsky S, Ott R, Silver N, Tidor B, Ravetch J, Wittrup K . Aglycosylated immunoglobulin G1 variants productively engage activating Fc receptors. Proc Natl Acad Sci U S A. 2008; 105(51):20167-72. PMC: 2629253. DOI: 10.1073/pnas.0809257105. View

Powell A, Harvey D . Stabilization of sialic acids in N-linked oligosaccharides and gangliosides for analysis by positive ion matrix-assisted laser desorption/ionization mass spectrometry. Rapid Commun Mass Spectrom. 1996; 10(9):1027-32. DOI: 10.1002/(SICI)1097-0231(19960715)10:9<1027::AID-RCM634>3.0.CO;2-Y. View

Hosokawa N, Tremblay L, Sleno B, Kamiya Y, Wada I, Nagata K . EDEM1 accelerates the trimming of alpha1,2-linked mannose on the C branch of N-glycans. Glycobiology. 2010; 20(5):567-75. DOI: 10.1093/glycob/cwq001. View

Stanley P . Golgi glycosylation. Cold Spring Harb Perspect Biol. 2011; 3(4). PMC: 3062213. DOI: 10.1101/cshperspect.a005199. View

Kelleher D, Gilmore R . An evolving view of the eukaryotic oligosaccharyltransferase. Glycobiology. 2005; 16(4):47R-62R. DOI: 10.1093/glycob/cwj066. View

10.

Hebert D, Garman S, Molinari M . The glycan code of the endoplasmic reticulum: asparagine-linked carbohydrates as protein maturation and quality-control tags. Trends Cell Biol. 2005; 15(7):364-70. DOI: 10.1016/j.tcb.2005.05.007. View

11.

Raykhel I, Alanen H, Salo K, Jurvansuu J, Nguyen V, Latva-Ranta M . A molecular specificity code for the three mammalian KDEL receptors. J Cell Biol. 2007; 179(6):1193-204. PMC: 2140024. DOI: 10.1083/jcb.200705180. View

12.

Faid V, Evjen G, Tollersrud O, Michalski J, Morelle W . Site-specific glycosylation analysis of the bovine lysosomal alpha-mannosidase. Glycobiology. 2006; 16(5):440-61. DOI: 10.1093/glycob/cwj081. View

13.

Trimble R, Maley F, Chu F . GlycoProtein biosynthesis in yeast. protein conformation affects processing of high mannose oligosaccharides on carboxypeptidase Y and invertase. J Biol Chem. 1983; 258(4):2562-7. View

14.

Kemmink J, Darby N, Dijkstra K, Nilges M, Creighton T . Structure determination of the N-terminal thioredoxin-like domain of protein disulfide isomerase using multidimensional heteronuclear 13C/15N NMR spectroscopy. Biochemistry. 1996; 35(24):7684-91. DOI: 10.1021/bi960335m. View

15.

Shental-Bechor D, Levy Y . Folding of glycoproteins: toward understanding the biophysics of the glycosylation code. Curr Opin Struct Biol. 2009; 19(5):524-33. DOI: 10.1016/j.sbi.2009.07.002. View

16.

Dell A, Galadari A, Sastre F, Hitchen P . Similarities and differences in the glycosylation mechanisms in prokaryotes and eukaryotes. Int J Microbiol. 2011; 2010:148178. PMC: 3068309. DOI: 10.1155/2010/148178. View

17.

Tian G, Xiang S, Noiva R, Lennarz W, Schindelin H . The crystal structure of yeast protein disulfide isomerase suggests cooperativity between its active sites. Cell. 2006; 124(1):61-73. DOI: 10.1016/j.cell.2005.10.044. View

18.

Geisler C, Jarvis D . Substrate specificities and intracellular distributions of three N-glycan processing enzymes functioning at a key branch point in the insect N-glycosylation pathway. J Biol Chem. 2012; 287(10):7084-97. PMC: 3293541. DOI: 10.1074/jbc.M111.296814. View

19.

Domon B, Vath J, Costello C . Analysis of derivatized ceramides and neutral glycosphingolipids by high-performance tandem mass spectrometry. Anal Biochem. 1990; 184(1):151-64. DOI: 10.1016/0003-2697(90)90028-8. View

20.

SCHACHTER H . The 'yellow brick road' to branched complex N-glycans. Glycobiology. 1991; 1(5):453-61. DOI: 10.1093/glycob/1.5.453. View