Jump to ContentJump to Main Navigation
Show Summary Details

Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Sies, Helmut / Turk, Boris / Wittinghofer, Alfred

12 Issues per year


IMPACT FACTOR 2015: 2.710
Rank 142 out of 289 in category Biochemistry & Molecular Biology in the 2015 Thomson Reuters Journal Citation Report/Science Edition

SCImago Journal Rank (SJR) 2015: 1.607
Source Normalized Impact per Paper (SNIP) 2015: 0.751
Impact per Publication (IPP) 2015: 2.609

Online
ISSN
1437-4315
See all formats and pricing
Volume 382, Issue 2 (Feb 2001)

Issues

Biosynthesis of Lipid-Linked Oligosaccharides in Yeast: the ALG3 Gene Encodes the Dol-P-Man:Man5GlcNAc2-PP-Dol Mannosyltransferase

Chandra B. Sharma / Roland Knauer / Ludwig Lehle
Published Online: 2005-06-01 | DOI: https://doi.org/10.1515/BC.2001.039

Abstract

The formation of Nglycosidic linkages of glycoproteins involves the ordered assembly of the common Glc[3]Man[9]GlcNAc[2] coreoligosaccharide on the lipid carrier dolichyl pyrophosphate. Whereas early mannosylation steps occur on the cytoplasmic side of the endoplasmic reticulum with GDPMan as donor, the final reactions from Man[5]GlcNAc[2]PPDol to Man[9]Glc Nac[2]PPDol on the lumenal side use DolPMan. We have investigated these later stages in vitro using a detergentsolubilized enzyme extract from yeast membranes. Mannosyltransfer from DolPMan to [[3]H]Man[5]GlcNAc[2]PPDol with formation of all intermediates up to Man[9]GlcNAc[2]PPDol occured in a rapid, time and proteindependent fashion. We find that the initial reaction from Man[5]GlcNAc[2]PPDol to Man[6]GlcNAc[2]PPDol is independent of metal ions, but further elongations need Mn[2+] that can be partly replaced by Mg[2+] or Ca[2+]. Zn[2+] or Cd[2+] ions were found to inhibit formation of Man[7-9]GlcNAc[2]PPDol, but do not affect synthesis of Man[6]GlcNAc[2]PPDol. Extension did not occur when the acceptor was added as a free Man[5]GlcNAc[2] oligosaccharide or when GDPMan was used as mannosyldonor. The alg3 mutant was described to accumulate Man[5]GlcNAc[2]PPDol. We expressed a functional active HAepitope tagged ALG3 fusion and succeeded to selectively immunoprecipitate the DolPMan:Man[5]GlcNAc[2]PPDol mannosyltransferase activity from the other enzymes of the detergent extract involved in the subsequent mannosylation reactions. This demonstrates that Alg3p represents the mannosyltransferase itself and not an accessory protein involved in the reaction.

About the article

Published Online: 2005-06-01

Published in Print: 2001-02-12


Citation Information: Biological Chemistry, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.2001.039. Export Citation

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
Shannon K. Esher, Joshua A. Granek, and J. Andrew Alspaugh
Fungal Genetics and Biology, 2015
[2]
E. Berends, L. Lehle, M. Henquet, T. Hesselink, H. A. Wosten, L. G. Lugones, and D. Bosch
Glycobiology, 2013, Volume 23, Number 2, Page 147
[3]
Widmar Tanner and Ludwig Lehle
Biochemical and Biophysical Research Communications, 2012, Volume 425, Number 3, Page 578
[4]
R. Oriol, I. Martinez-Duncker, I. Chantret, R. Mollicone, and P. Codogno
Molecular Biology and Evolution, 2002, Volume 19, Number 9, Page 1451
[5]
Toshihide Nakamura, Akira Ando, Hiroshi Takagi, and Jun Shima
Biochemical and Biophysical Research Communications, 2007, Volume 353, Number 2, Page 293
[6]
Anna Janik, Monika Sosnowska, Joanna Kruszewska, Hubert Krotkiewski, Ludwig Lehle, and Grażyna Palamarczyk
Biochimica et Biophysica Acta (BBA) - General Subjects, 2003, Volume 1621, Number 1, Page 22
[7]
Juergen H. Nett, Terrance A. Stadheim, Huijuan Li, Piotr Bobrowicz, Stephen R. Hamilton, Robert C. Davidson, Byung-Kwon Choi, Teresa Mitchell, Beata Bobrowicz, Alissa Rittenhour, Stefan Wildt, and Tillman U. Gerngross
Yeast, 2011, Volume 28, Number 3, Page 237
[8]
Masahiko Miyamoto, Yasuhiro Furuichi, and Tadazumi Komiyama
Yeast, 2011, Volume 28, Number 1, Page 27
[10]
K. Hese, C. Otto, F. H Routier, and L. Lehle
Glycobiology, 2008, Volume 19, Number 2, Page 160
[11]
Stefan Wildt and Tilllman U. Gerngross
Nature Reviews Microbiology, 2005, Volume 3, Number 2, Page 119
[12]
Yeast, 2001, Volume 18, Number 11, Page 1091

Comments (0)

Please log in or register to comment.
Log in