Abstract
An alternativeN-glycosylation pathway using Glc1–3Man5GlcNAc2 as a donor to be transferred to a protein acceptor is found either in Man-P-Dol synthase deficient cells or in wild type CHO cells grown in energy deprivation conditions. Discrimination between oligomannosides of this alternative pathway and oligomannosides of the major one containing the same number of sugar residues Man6–8GlcNAc2 required structural studies. Taking advantage of the specific chromatographic behaviour of glucosylated oligomannosides, in pellicular high pH anion exchange chromatography, we developed a one-step method for the identification of the alternativeN-glycosylation pathway compounds differing from those of the major one.
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Abbreviations
- HPAEC:
-
high pH anion exchange chromatography
- endo H:
-
endo betaN-acetylglucosaminidase H
- PNGaseF:
-
peptideN-glycosidase F
- M2:
-
Man2GlcNAc2
- M4:
-
Man4GlcNAc2
- M5:
-
Man5GlcNAc2
- G1M5:
-
Glc1Man5GlcNAc2
- G2M5:
-
Glc2Man5GlcNAc2
- G3M5:
-
Glc3Man5GlcNAc2
- M6:
-
Man6GlcNAc2
- M8:
-
Man8GlcNAc2
- M9:
-
Man9GlcNAc2
- G1M9:
-
Glc1Man9GlcNAc2
- G2M9:
-
Glc2Man9GlcNAc2
- G3M9:
-
Glc3Man9GlcNAc2
References
Chapman A, Fujimoto K, Kornfeld S (1980)J Biol Chem 255:4441–46.
Stoll J, Krag SS (1983)J Cell Biol 97:443a.
Stoll J (1986) PhD Thesis. School of Hygiene and Public Health, The Johns Hopkins University, Baltimore, Maryland, USA.
Turco SJ (1981)Anal Biochem 118:278–83.
Turco SJ, Stetson B, Robbins PW (1977)Proc Natl Acad Sci USA 74:4411–14.
Hernandez LM, Ballou L, Ballou CE (1990)Carbohydr Res 203:1–11.
Hermentin P, Witzel R, Vliegenthart JFG, Kamerling JP, Nimtz M, Conradt HS (1991)Anal Biochem 203:281–89.
Nimtz M, Martin W, Wray V, Kloppel KD, Augustin J, Conradt HS (1993)Eur J Biochem 213:39–56.
Cacan R, Villers C, Bélard M, Kaiden A, Krag S, Verbert A (1992)Glycobiology 2:127–36.
Rearick JI, Chapman A, Kornfeld S (1981)J Biol Chem 256:6255–61.
Chapman AE, Calhoun JC (1988)Arch Bioch Biophys 260:320–33.
Maley F, Trimble RB, Tarentino AL, Plummer TS, Jr (1989)Anal Biochem 180:195–204.
Alvarado E, Nukada T, Ogawa T, Ballou CE (1991)Biochemistry 30:881–86.
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Villers, C., Plancke, Y., Cacan, R. et al. Rapid detection of the alternativeN-glycosylation pathway using high pH anion exchange chromatography. Glycoconjugate J 11, 226–231 (1994). https://doi.org/10.1007/BF00731222
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DOI: https://doi.org/10.1007/BF00731222