Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred

IMPACT FACTOR 2018: 3.014
5-year IMPACT FACTOR: 3.162

CiteScore 2018: 3.09

SCImago Journal Rank (SJR) 2018: 1.482
Source Normalized Impact per Paper (SNIP) 2018: 0.820

See all formats and pricing
More options …
Volume 382, Issue 2


Cloning and Expression of Drosophila melanogaster UDP-GlcNAc:?-3-D-Mannoside ? 1,2-N-Acetylglucosaminyltransferase I

Mohan Sarkar / Harry Schachter
Published Online: 2005-06-01 | DOI: https://doi.org/10.1515/BC.2001.028


A TBLASTN search of the Drosophila melanogaster expressed sequence tag (EST) database with the amino acid sequence of human UDPNacetylglucosamine: ?-3-Dmannoside ?-1,2-Nacetylglucosaminyltransferase I (GnT I, EC as probe yielded a clone (GM01211) with 56% identity over 36 carboxyterminal amino acids. A 550 base pair (bp) probe derived from the EST clone was used to screen a Drosophila cDNA library in ?ZAP II and two cDNAs lacking a start ATG codon were obtained. 5Rapid amplification of cDNA ends (5RACE) yielded a 2828 bp cDNA containing a fulllength 1368 bp open reading frame encoding a 456 amino acid protein with putative Nterminal cytoplasmic (5 residues) and hydrophobic transmembrane (20 residues) domains. The protein showed 52% amino acid sequence identity to human GnT I. This cDNA, truncated to remove the Nterminal hydrophobic domain, was expressed in the baculovirus/Sf9 system as a secreted protein containing an Nterminal (His)[6] tag. Protein purified by adsorption to and elution from nickel beads converted Man?1-6(Man?1-3)Man?octyl (M3-octyl) to Man?1-6(GlcNAc?1-2Man?1-3)Man?octyl. The K values (0.7 and 0.03 mM for M3-octyl and UDPGlc NAc respectively), temperature optimum (37 C), pH optimum (pH 5 to 6) and divalent cation requirements (Mn > Fe, Mg, Ni > Ba, Ca, Cd, Cu) were similar to mammalian GnT I. TBLASTN searches of the Berkeley Drosophila Genome Project database with the Drosophila GnT I cDNA sequence as probe allowed localization of the gene to chromosomal region 2R; 57A9. Comparison of the cDNA and genomic DNA sequences allowed the assignment of seven exons and six introns; all introns showed GTAG splice site consensus sequences. This is the first insect GnT I gene to be cloned and expressed.

About the article

Published Online: 2005-06-01

Published in Print: 2001-02-12

Citation Information: Biological Chemistry, Volume 382, Issue 2, Pages 209–217, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.2001.028.

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.

Sarah P. Lawson, Leah T. Sigle, Abigail L. Lind, Andrew W. Legan, Jessica N. Mezzanotte, Hans-Willi Honegger, and Patrick Abbot
Evolution, 2017, Volume 71, Number 8, Page 1986
Christoph Geisler, Varshika Kotu, Mary Sharrow, Dubravko Rendić, Gerald Pöltl, Michael Tiemeyer, Iain B. H. Wilson, and Donald L. Jarvis
Journal of Biological Chemistry, 2012, Volume 287, Number 35, Page 29599
Renaud Léonard, Dubravko Rendić, Catherine Rabouille, Iain B. H. Wilson, Thomas Préat, and Friedrich Altmann
Journal of Biological Chemistry, 2006, Volume 281, Number 8, Page 4867
Dubravko Rendić, Angela Linder, Katharina Paschinger, Nicole Borth, Iain B. H. Wilson, and Gustáv Fabini
Journal of Biological Chemistry, 2006, Volume 281, Number 6, Page 3343
Kazuhiro Aoki, Mindy Perlman, Jae-Min Lim, Rebecca Cantu, Lance Wells, and Michael Tiemeyer
Journal of Biological Chemistry, 2007, Volume 282, Number 12, Page 9127
Gustáv Fabini, Angelika Freilinger, Friedrich Altmann, and Iain B. H. Wilson
Journal of Biological Chemistry, 2001, Volume 276, Number 30, Page 28058
Mohan Sarkar, Peter A. Leventis, Cristina I. Silvescu, Vernon N. Reinhold, Harry Schachter, and Gabrielle L. Boulianne
Journal of Biological Chemistry, 2006, Volume 281, Number 18, Page 12776
Tomomi Ichimiya, Hiroshi Manya, Yoshiko Ohmae, Hideki Yoshida, Kuniaki Takahashi, Ryu Ueda, Tamao Endo, and Shoko Nishihara
Journal of Biological Chemistry, 2004, Volume 279, Number 41, Page 42638
Vladimir Sladek and Igor Tvaroška
The Journal of Physical Chemistry B, 2017, Volume 121, Number 25, Page 6148
Rhiannon Stanton, Alba Hykollari, Barbara Eckmair, Daniel Malzl, Martin Dragosits, Dieter Palmberger, Ping Wang, Iain B.H. Wilson, and Katharina Paschinger
Biochimica et Biophysica Acta (BBA) - General Subjects, 2017, Volume 1861, Number 4, Page 699
Nathan T. Mortimer, Balint Z. Kacsoh, Erin S. Keebaugh, Todd A. Schlenke, and David S. Schneider
PLoS Pathogens, 2012, Volume 8, Number 7, Page e1002819
Hideaki Mabashi-Asazuma, Chu-Wei Kuo, Kay-Hooi Khoo, and Donald L. Jarvis
ACS Chemical Biology, 2015, Volume 10, Number 10, Page 2199
Miki Yamamoto-Hino, Hideki Yoshida, Tomomi Ichimiya, Sho Sakamura, Megumi Maeda, Yoshinobu Kimura, Norihiko Sasaki, Kiyoko F. Aoki-Kinoshita, Akiko Kinoshita-Toyoda, Hidenao Toyoda, Ryu Ueda, Shoko Nishihara, and Satoshi Goto
Genes to Cells, 2015, Volume 20, Number 6, Page 521
Harry Schachter and Gabrielle Boulianne
Fly, 2011, Volume 5, Number 1, Page 18
Miki Yamamoto-Hino, Masato Abe, Takako Shibano, Yuka Setoguchi, Wakae Awano, Ryu Ueda, Hideyuki Okano, and Satoshi Goto
Cell Structure and Function, 2012, Volume 37, Number 1, Page 55
Koji Dohi, Junko Isoyama-Tanaka, Toru Tokuda, and Kazuhito Fujiyama
Journal of Bioscience and Bioengineering, 2010, Volume 109, Number 4, Page 388
Harry Schachter
Carbohydrate Research, 2009, Volume 344, Number 12, Page 1391
Sonia G Tsitilou and Sofia Grammenoudi
Biochemical and Biophysical Research Communications, 2003, Volume 312, Number 4, Page 1372
Shihao Chen, Jenny Tan, Vernon N Reinhold, Andrew M Spence, and Harry Schachter
Biochimica et Biophysica Acta (BBA) - General Subjects, 2002, Volume 1573, Number 3, Page 271
Friedrich Altmann, Gustáv Fabini, Horst Ahorn, and Iain B.H. Wilson
Biochimie, 2001, Volume 83, Number 8, Page 703
M. Sarkar, K. G. Iliadi, P. A. Leventis, H. Schachter, and G. L. Boulianne
Proceedings of the National Academy of Sciences, 2010, Volume 107, Number 21, Page 9677
Robert S. Haltiwanger and John B. Lowe
Annual Review of Biochemistry, 2004, Volume 73, Number 1, Page 491
T. Okada, H. Ihara, R. Ito, M. Nakano, K. Matsumoto, Y. Yamaguchi, N. Taniguchi, and Y. Ikeda
Glycobiology, 2010, Volume 20, Number 9, Page 1147
Noboru Tomiya
Trends in Glycoscience and Glycotechnology, 2009, Volume 21, Number 118, Page 71
Dubravko Rendić, Iain B. H. Wilson, Gert Lubec, Martin Gutternigg, Friedrich Altmann, and Renaud Léonard
ELECTROPHORESIS, 2007, Volume 28, Number 23, Page 4484
Yeon Kyu Kim, Hwa Sung Shin, Noboru Tomiya, Yuan C. Lee, Michael J. Betenbaugh, and Hyung Joon Cha
Biotechnology and Bioengineering, 2005, Volume 92, Number 4, Page 452
Renaud L�onard, Daniel Kolarich, Katharina Paschinger, Friedrich Altmann, and Iain Wilson
Plant Molecular Biology, 2004, Volume 55, Number 5, Page 631

Comments (0)

Please log in or register to comment.
Log in