Oligosaccharyltransferase

Oligosaccharyltransferase or OST (EC 2.4.1.119) is a membrane protein complex that transfers a 14-sugar oligosaccharide from dolichol to nascent protein. It is a type of glycosyltransferase. The sugar Glc₃Man₉GlcNAc₂ (where Glc=Glucose, Man=Mannose, and GlcNAc=N-acetylglucosamine) is attached to an asparagine (Asn) residue in the sequence Asn-X-Ser or Asn-X-Thr where X is any amino acid except proline. This sequence is called a glycosylation sequon. The reaction catalyzed by OST is the central step in the N-linked glycosylation pathway.

STT3
Structure of yeast OST [1]
Identifiers
Symbol	STT3
Pfam	PF02516
Pfam clan	CL0111
InterPro	IPR003674
OPM superfamily	242
OPM protein	3rce
CAZy	GT66
Membranome	275
Available protein structures: Pfam   structures / ECOD   PDB RCSB PDB; PDBe; PDBj PDBsum structure summary

Location

ER Translocon complex.^[2] Many protein complexes are involved in protein synthesis. The actual production takes place in the ribosomes (grey and light blue). Through the ER translocon (green: Sec61, blue: TRAP complex, and red: oligosaccharyl transferase complex) the newly synthesized protein is transported across the membrane (gray) into the interior of the ER. Sec61 is the protein-conducting channel and the OST adds sugar moieties to the nascent protein.

OST is a component of the translocon in the endoplasmic reticulum (ER) membrane. A lipid-linked core-oligosaccharide is assembled at the membrane of the endoplasmic reticulum and transferred to selected asparagine residues of nascent polypeptide chains by the oligosaccharyl transferase complex.^[3] The active site of OST is located about 4 nm from the lumenal face of the ER membrane.^[4]

It usually acts during translation as the nascent protein is entering the ER, but this cotranslational glycosylation is nevertheless called a posttranslational modification. A few examples have been found of OST activity after translation is complete.^[5][6] Current opinion is that post-translational activity may occur if the protein is poorly folded or folds slowly.^[6]

Structure and function

Yeast OST is composed of eight different membrane-spanning proteins in three subcomplexes (one of them is OST4).^[7][8] These octomers do not form higher order oligomers, and three of the eight proteins are glycosylated themselves.^[7] OST in mammals is known to have a similar composition.^[9][10]

OST is thought to require many subunits because it must:^[11]

1. Position itself near the translocon pore.
2. Recognize and bind oligosaccharyldolichol.
3. Scan the nascent protein in order to recognize and bind sequons.
4. Move these two large substrates into their proper locations and conformations.
5. Activate the Asn amide nitrogen atom for the actual transfer of oligosaccharide.
6. Release its substrates.

The catalytically active subunit of the OST is called STT3. Two paralogs exist in eukaryotes, termed STT3A and STT3B. STT3A is primarily responsible for cotranslational glycosylation of the nascent polypeptide as it enters the lumen of the endoplasmic reticulum whereas STT3B can also mediate posttranslational glycosylation.^[12] The structure of PglB, the prokaryotic homolog of STT3 has been solved.^[13] The high sequence similarity between the prokaryotic and the eukaryotic STT3 suggests that their structures are similar.

Clinical significance

CDG syndromes are genetic disorders of the glycosylation pathway. They are labelled "Type I" if the defective gene is for an enzyme involved in the assembly or transfer of the Glc₃Man₉GlcNAc₂-dolichol precursor. They are labelled "Type II" if the defective step occurs after the action of OST in the N-linked glycosylation pathway or involves O-linked glycosylation.^[14]

See also

• STT3A
• STT3B