Home LiteratureArticle Details
PMID: 20007322 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Comparative structural biology of eubacterial and archaeal oligosaccharyltransferases.

The Journal of biological chemistry ·Vol. 285 ·No. 7 ·2010-02-12 ·Pages 4941-50

Maita N, Nyirenda J, Igura M, Kamishikiryo J, Kohda D

Abstract

Oligosaccharyltransferase (OST) catalyzes the transfer of an oligosaccharide from a lipid donor to an asparagine residue in nascent polypeptide chains. In the bacterium Campylobacter jejuni, a single-subunit membrane protein, PglB, catalyzes N-glycosylation. We report the 2.8 A resolution crystal structure of the C-terminal globular domain of PglB and its comparison with the previously determined structure from the archaeon Pyrococcus AglB. The two distantly related oligosaccharyltransferases share unexpected structural similarity beyond that expected from the sequence comparison. The common architecture of the putative catalytic sites revealed a new catalytic motif in PglB. Site-directed mutagenesis analyses confirmed the contribution of this motif to the catalytic function. Bacterial PglB and archaeal AglB constitute a protein family of the catalytic subunit of OST along with STT3 from eukaryotes. A structure-aided multiple sequence alignment of the STT3/PglB/AglB protein family revealed three types of OST catalytic centers. This novel classification will provide a useful framework for understanding the enzymatic properties of the OST enzymes from Eukarya, Archaea, and Bacteria.

MeSH Terms
Archaeal Proteins/chemistry,genetics,metabolism Bacterial Proteins/chemistry,genetics,metabolism Campylobacter jejuni/enzymology Crystallography, X-Ray Hexosyltransferases/chemistry,genetics,metabolism Membrane Proteins/chemistry,genetics,metabolism Models, Biological Protein Structure, Secondary Protein Structure, Tertiary/genetics,physiology Pyrococcus/enzymology
Chemicals
Archaeal Proteins Bacterial Proteins Membrane Proteins Hexosyltransferases dolichyl-diphosphooligosaccharide - protein glycotransferase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Maita Nobuo
Division of Structural Biology, Medical Institute of Bioregulation, Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka 812-8582, Japan.
Nyirenda James
Igura Mayumi
Kamishikiryo Jun
Kohda Daisuke
References (44)
44 references, click to expand
  1. Distinct donor and acceptor specificities of Trypanosoma brucei oligosaccharyltransferases.
    EMBO J. 2009 Sep 2;28(17):2650-61 PMID: 19629045
  2. Protein glycosylation in bacterial mucosal pathogens.
    Nat Rev Microbiol. 2005 Mar;3(3):225-37 PMID: 15738950
  3. Oligosaccharyltransferase isoforms that contain different catalytic STT3 subunits have distinct enzymatic properties.
    Mol Cell. 2003 Jul;12(1):101-11 PMID: 12887896
  4. New oligosaccharyltransferase assay method.
    Glycobiology. 2007 Nov;17(11):1175-82 PMID: 17693440
  5. Yeast oligosaccharyltransferase consists of two functionally distinct sub-complexes, specified by either the Ost3p or Ost6p subunit.
    FEBS Lett. 2005 Dec 5;579(29):6564-8 PMID: 16297388
  6. Ribophorin I regulates substrate delivery to the oligosaccharyltransferase core.
    Proc Natl Acad Sci U S A. 2008 Jul 15;105(28):9534-9 PMID: 18607003
  7. From peptide to protein: comparative analysis of the substrate specificity of N-linked glycosylation in C. jejuni.
    Biochemistry. 2007 May 8;46(18):5579-85 PMID: 17439157
  8. Studies on the function of oligosaccharyl transferase subunits. Stt3p is directly involved in the glycosylation process.
    J Biol Chem. 2002 Dec 6;277(49):47692-700 PMID: 12359722
  9. The oligosaccharyltransferase complex from yeast.
    Biochim Biophys Acta. 1999 Jan 6;1426(2):259-73 PMID: 9878773
  10. The CCP4 suite: programs for protein crystallography.
    Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3 PMID: 15299374
  11. The yeast oligosaccharyltransferase complex can be replaced by STT3 from Leishmania major.
    Glycobiology. 2009 Feb;19(2):160-71 PMID: 18955371
  12. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform.
    Nucleic Acids Res. 2002 Jul 15;30(14):3059-66 PMID: 12136088
  13. The N-terminal carbohydrate recognition site of the cation-independent mannose 6-phosphate receptor.
    J Biol Chem. 2004 Aug 6;279(32):34000-9 PMID: 15169779
  14. Structure of the oligosaccharyl transferase complex at 12 A resolution.
    Structure. 2008 Mar;16(3):432-40 PMID: 18334218
  15. Identification of genes involved in the biosynthesis and attachment of Methanococcus voltae N-linked glycans: insight into N-linked glycosylation pathways in Archaea.
    Mol Microbiol. 2006 Jul;61(1):259-68 PMID: 16824110
  16. All in one: Leishmania major STT3 proteins substitute for the whole oligosaccharyltransferase complex in Saccharomyces cerevisiae.
    Mol Biol Cell. 2008 Sep;19(9):3758-68 PMID: 18596231
  17. Structure-guided identification of a new catalytic motif of oligosaccharyltransferase.
    EMBO J. 2008 Jan 9;27(1):234-43 PMID: 18046457
  18. N-linked glycosylation in Campylobacter jejuni and its functional transfer into E. coli.
    Science. 2002 Nov 29;298(5599):1790-3 PMID: 12459590
  19. Unraveling the mechanism of protein N-glycosylation.
    J Biol Chem. 2005 Feb 4;280(5):3121-4 PMID: 15590627
  20. Crystallography & NMR system: A new software suite for macromolecular structure determination.
    Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21 PMID: 9757107
  21. Facing extremes: archaeal surface-layer (glyco)proteins.
    Microbiology (Reading). 2003 Dec;149(Pt 12):3347-3351 PMID: 14663068
  22. Preferential transfer of the complete glycan is determined by the oligosaccharyltransferase complex and not by the catalytic subunit.
    Proc Natl Acad Sci U S A. 2006 Oct 3;103(40):14756-60 PMID: 17001015
  23. An analysis of amino acid sequences surrounding archaeal glycoprotein sequons.
    Archaea. 2007 May;2(2):73-81 PMID: 17350928
  24. Processing of X-ray diffraction data collected in oscillation mode.
    Methods Enzymol. 1997;276:307-26 PMID: 27754618
  25. Cotranslational and posttranslational N-glycosylation of polypeptides by distinct mammalian OST isoforms.
    Cell. 2009 Jan 23;136(2):272-83 PMID: 19167329
  26. Dolichol-linked oligosaccharide selection by the oligosaccharyltransferase in protist and fungal organisms.
    J Cell Biol. 2007 Apr 9;177(1):29-37 PMID: 17403929
  27. Lysine methylation as a routine rescue strategy for protein crystallization.
    Structure. 2006 Nov;14(11):1617-22 PMID: 17098187
  28. Engineering N-linked protein glycosylation with diverse O antigen lipopolysaccharide structures in Escherichia coli.
    Proc Natl Acad Sci U S A. 2005 Feb 22;102(8):3016-21 PMID: 15703289
  29. Membrane topology of the STT3 subunit of the oligosaccharyl transferase complex.
    J Biol Chem. 2005 May 27;280(21):20261-7 PMID: 15781470
  30. Identification and characterization of the unique N-linked glycan common to the flagellins and S-layer glycoprotein of Methanococcus voltae.
    J Biol Chem. 2005 Apr 29;280(17):16586-93 PMID: 15723834
  31. Protein glycosylation: nature, distribution, enzymatic formation, and disease implications of glycopeptide bonds.
    Glycobiology. 2002 Apr;12(4):43R-56R PMID: 12042244
  32. The STT3a subunit isoform of the Arabidopsis oligosaccharyltransferase controls adaptive responses to salt/osmotic stress.
    Plant Cell. 2003 Oct;15(10):2273-84 PMID: 12972670
  33. Dimeric organization of the yeast oligosaccharyl transferase complex.
    Proc Natl Acad Sci U S A. 2006 Jun 13;103(24):8947-52 PMID: 16754853
  34. Coot: model-building tools for molecular graphics.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32 PMID: 15572765
  35. Purification, crystallization and preliminary X-ray diffraction studies of the soluble domain of the oligosaccharyltransferase STT3 subunit from the thermophilic archaeon Pyrococcus furiosus.
    Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Sep 1;63(Pt 9):798-801 PMID: 17768359
  36. An evolving view of the eukaryotic oligosaccharyltransferase.
    Glycobiology. 2006 Apr;16(4):47R-62R PMID: 16317064
  37. Oxidoreductase activity of oligosaccharyltransferase subunits Ost3p and Ost6p defines site-specific glycosylation efficiency.
    Proc Natl Acad Sci U S A. 2009 Jul 7;106(27):11061-6 PMID: 19549845
  38. Automated structure solution with autoSHARP.
    Methods Mol Biol. 2007;364:215-30 PMID: 17172768
  39. GASH: an improved algorithm for maximizing the number of equivalent residues between two protein structures.
    BMC Bioinformatics. 2005 Sep 08;6:221 PMID: 16146579
  40. Deep-sea vent epsilon-proteobacterial genomes provide insights into emergence of pathogens.
    Proc Natl Acad Sci U S A. 2007 Jul 17;104(29):12146-50 PMID: 17615243
  41. Chemoenzymatic synthesis of glycopeptides with PglB, a bacterial oligosaccharyl transferase from Campylobacter jejuni.
    Chem Biol. 2005 Dec;12(12):1311-5 PMID: 16356848
  42. Three monophyletic superfamilies account for the majority of the known glycosyltransferases.
    Protein Sci. 2003 Jul;12(7):1418-31 PMID: 12824488
  43. Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering.
    Protein Eng. 1990 Apr;3(5):433-42 PMID: 2349213
  44. Ribophorin I acts as a substrate-specific facilitator of N-glycosylation.
    J Cell Sci. 2007 Feb 15;120(Pt 4):648-57 PMID: 17264154
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M109.081752
Published
2010-02-12
Epub
2009-00-09
Pages
4941-50
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2836098
Subset
IM
Databases
PDB
3AAG
External Links
PubMed source DOI: 10.1074/jbc.M109.081752 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2