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Post-Translational Events II. ER & Golgi Processes. Rough Endoplasmic Reticulum. Glycolsylation. O-linked – serine, threonine, hydroxylysine (collagen) N-linked - asparagine The sugars GalNac – N-acetylgalactosamine GlcNac – N-acetylglucosamine NANA – N-acetylneuraminic acid (sialic acid)
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Post-Translational Events II ER & Golgi Processes
Glycolsylation • O-linked – serine, threonine, hydroxylysine (collagen) • N-linked - asparagine • The sugars • GalNac – N-acetylgalactosamine • GlcNac – N-acetylglucosamine • NANA – N-acetylneuraminic acid (sialic acid) • Gal – galactose • Glc – glucose • Man – mannose • Fuc – fucose • Dolicholpyrophosphoryl oligosaccharide • All sugars are linked to nucleoside mono- or diphosphate • UDP-gal, UDP-GalNac, UDP-GlcNac • GDP-man • CMP-NANA • Dolichol pyrophosphate
Glyco-nucleoside Phosphates CH2CH(CH3)CH2CH2OPO3 CH3C(CH3)=CHCH2 CH2C(CH3)=CHCH2 15-18 Dolichol phosphate
O-linked Glycosylation • Small chains of sugars • N-acetylgalactosamine (GalNac) is first sugar on S orT • GalNac transferase in ER and cis-golgi • Additional sugars added in cis, medial and trans-golgi • Gal added by galactosyl transferase • NANA added last • Each sugar addition is catalyzed by specific enzyme
N-linked Glycosylation • Initial step in ER • Dolichol pyrophosphoryl oligosaccharide • 14 sugar moiety attached to dolichol pyrophosphate • (GalNac)2Man9Glc3 • Dolichol phosphate in ER membrane • Sugars added by cytostolic glycosyltransferases • Moiety flips in ER membrane to face lumen • Transferred to asparagine by oligosaccharyl transferase (oligosaccharide-protein transferase) • Target sequence N-X-S and N-X-T (X = P) • Subsequent remodeling of sugar moiety in ER & golgi • N-linked chains very long and extensively branched
GPI – Linkages • GPI – linked proteins are attached to GPI in ER • GPI in ER membrane • C-term of protein attached to sugar residue on phosphatidyl inositol • GPI linked protein will be attached to outer leaflet of plasma membrane
Unique Signaling from ER to Make More ER • Pathway activated by stresses that result in more unfolded proteins • heat • unfavorable pHs
Processing in the Golgi • Oligosaccharide modifications • Sorting to exocytic vesicles, lysosomes and ER • Proteolytic processing
Modification of N-linked Oligosaccharides • As glycosylated protein moves through ER Glc is removed • In cis Golgi, some Man is removed • In medial golgi, GlcNac is added • In trans-golgi, Gal and NANA are added CIS TRANS
Sterol & Lipid Biosynthesis • Phospholgycerides in ER • Sphingolipids in Golgi • Lipid profile asymmetry established via phospholipid exchange proteins and selective vesicular transport due to association between carrier proteins and certain lipid types