Photoaffinity labeling of protein O-Mannosyltransferases of the PMT1/PMT2 subfamily

Protein O-mannosylation is initiated at the endoplasmic reticulum (ER) by dolichyl phosphate-mannose: protein O-mannosyltransferases (PMTs). PMTs are members of the glycosyltransferase (GT) C superfamily. They are large polytopic integral membrane proteins located in the ER membrane. PMTs utilize do...

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Hauptverfasser: Loibl, Martin (VerfasserIn) , Strahl, Sabine (VerfasserIn)
Dokumenttyp: Kapitel/Artikel
Sprache:Englisch
Veröffentlicht: 16 May 2013
In: Glycosyltransferases
Year: 2013, Pages: 107-117
DOI:10.1007/978-1-62703-465-4_9
Online-Zugang:Verlag, Volltext: http://dx.doi.org/10.1007/978-1-62703-465-4_9
Verlag, Volltext: http:/http://link.springer.com/protocol/10.1007%2F978-1-62703-465-4_9
Volltext
Verfasserangaben:Martin Loibl, Sabine Strahl

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520 |a Protein O-mannosylation is initiated at the endoplasmic reticulum (ER) by dolichyl phosphate-mannose: protein O-mannosyltransferases (PMTs). PMTs are members of the glycosyltransferase (GT) C superfamily. They are large polytopic integral membrane proteins located in the ER membrane. PMTs utilize dolichyl phosphate-­activated mannose as sugar donor. Glycosyltransfer of mannose to serine and threonine residues of nascent polypeptides leads to an inversion of the stereochemistry of the glycosidic bond. Here, we describe photoaffinity labeling of yeast Pmt1p using a photo-reactive probe that is based on the artificial mannosyl acceptor peptide YATAV. Due to the high homology of PMTs, this method can also be applied to study PMT1 and PMT2 subfamily members from fungi other than baker’s yeast. 
650 4 |a Cross-linking 
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