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Structure-guided identification of a new catalytic motif of oligosaccharyltransferase.

Igura M, Maita N, Kamishikiryo J, Yamada M, Obita T, Maenaka K, Kohda D
EMBO J 27 234-43 (2008)
Cited: 113 times
EuropePMC logo PMID: 18046457

Abstract

Asn-glycosylation is widespread not only in eukaryotes but also in archaea and some eubacteria. Oligosaccharyltransferase (OST) catalyzes the co-translational transfer of an oligosaccharide from a lipid donor to an asparagine residue in nascent polypeptide chains. Here, we report that a thermophilic archaeon, Pyrococcus furiosus OST is composed of the STT3 protein alone, and catalyzes the transfer of a heptasaccharide, containing one hexouronate and two pentose residues, onto peptides in an Asn-X-Thr/Ser-motif-dependent manner. We also determined the 2.7-A resolution crystal structure of the C-terminal soluble domain of Pyrococcus STT3. The structure-based multiple sequence alignment revealed a new motif, DxxK, which is adjacent to the well-conserved WWDYG motif in the tertiary structure. The mutagenesis of the DK motif residues in yeast STT3 revealed the essential role of the motif in the catalytic activity. The function of this motif may be related to the binding of the pyrophosphate group of lipid-linked oligosaccharide donors through a transiently bound cation. Our structure provides the first structural insights into the formation of the oligosaccharide-asparagine bond.

Articles - 2zai mentioned but not cited (4)

  1. Structure-guided identification of a new catalytic motif of oligosaccharyltransferase. Igura M, Maita N, Kamishikiryo J, Yamada M, Obita T, Maenaka K, Kohda D. EMBO J 27 234-243 (2008)
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  3. Rational design of crystal contact-free space in protein crystals for analyzing spatial distribution of motions within protein molecules. Matsuoka R, Shimada A, Komuro Y, Sugita Y, Kohda D. Protein Sci 25 754-768 (2016)
  4. Eukaryotic N-glycosylation occurs via the membrane-anchored C-terminal domain of the Stt3p subunit of oligosaccharyltransferase. Huang C, Bhaskaran R, Mohanty S. J Biol Chem 287 32450-32458 (2012)


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