Sequences from the first fibronectin type III repeat of the neural cell adhesion molecule allow O-glycan polysialylation of an adhesion molecule chimera.
Polysialic acid is a developmentally regulated, anti-adhesive polymer that is added to N-glycans on the fifth immunoglobulin domain (Ig5) of the neural cell adhesion molecule (NCAM). We found that the first fibronectin type III repeat (FN1) of NCAM is required for the polysialylation of N-glycans on the adjacent Ig5 domain, and we proposed that the polysialyltransferases recognize specific sequences in FN1 to position themselves for Ig5 N-glycan polysialylation. Other studies identified a novel FN1 acidic surface patch and α-helix that play roles in NCAM polysialylation. Here, we characterize the contribution of two additional FN1 sequences, Pro(510)-Tyr(511)-Ser(512) (PYS) and Gln(516)-Val(517)-Gln(518) (QVQ). Replacing PYS or the acidic patch dramatically decreases the O-glycan polysialylation of a truncated NCAM protein, and replacing the α-helix or QVQ shifts polysialic acid to FN1 O-glycans in full-length NCAM. We also found that the FN1 domain of the olfactory cell adhesion molecule, a homologous but unpolysialylated protein, could partially replace NCAM FN1. Inserting Pro(510)-Tyr(511) eliminated N-glycan polysialylation and enhanced O-glycosylation of an NCAM- olfactory cell adhesion molecule chimera, and inserting other FN1 sequences unique to NCAM, predominantly the acidic patch, created a new polysialyltransferase recognition site. Taken together, our results highlight the role of the FN1 α-helix and QVQ sequences in N-glycan polysialylation and demonstrate that the acidic patch primarily functions in O-glycan polysialylation.
Foley
Deirdre A
DA
Department of Biochemistry and Molecular Genetics, College of Medicine, University of Illinois, Chicago, Illinois 60607, USA.
Swartzentruber
Kristin G
KG
Thompson
Matthew G
MG
Mendiratta
Shalu Shiv
SS
Colley
Karen J
KJ
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D005353:Fibronectins; D006801:Humans; D019158:N-Acetylneuraminic Acid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019006:Neural Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism
D005353:Fibronectins; D019006:Neural Cell Adhesion Molecules; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; D012799:Sialyltransferases; D019158:N-Acetylneuraminic Acid
Mol Carcinog. 2010 Apr;49(4):386-97
20191608
Science. 1988 Apr 1;240(4848):53-7
3281256
Proc Natl Acad Sci U S A. 1988 May;85(9):2999-3003
2834727
J Biol Chem. 1992 May 15;267(14):9965-71
1315775
J Immunol. 2008 Nov 15;181(10):6850-8
18981104
J Biol Chem. 1996 Aug 9;271(32):19457-63
8702635
Glycobiology. 1996 Apr;6(3):289-301
8724137
Neurochem Res. 2004 Nov;29(11):2015-35
15662836
J Biol Chem. 2006 Nov 24;281(47):36052-9
17003032
J Biochem. 2006 Nov;140(5):687-701
17023684
J Biol Chem. 1997 Oct 17;272(42):26083-6
9334170
Lab Invest. 1994 Jan;70(1):95-106
8302024
Biochem Biophys Res Commun. 1983 Apr 29;112(2):482-7
6847662
J Neurosci. 1997 Aug 1;17(15):5830-42
9221781
J Biol Chem. 1995 Sep 29;270(39):22685-8
7559389
Brain Res Rev. 2007 Nov;56(1):101-18
17658613
J Cell Biol. 2009 Dec 28;187(7):1101-16
20038681
J Virol. 1989 Oct;63(10):4409-16
2778882
J Neurochem. 2005 Oct;95(2):570-83
16135080
Oncology. 2010;78(3-4):196-204
20414008
J Biol Chem. 2005 Sep 16;280(37):32340-8
16027151
Cancer Res. 1998 Feb 15;58(4):779-84
9485034
Glycobiology. 2005 Sep;15(9):887-94
15872150
Brain. 2009 Oct;132(Pt 10):2831-8
19443631
Science. 1987 May 15;236(4803):799-806
3576199
Proc Natl Acad Sci U S A. 2010 Jun 1;107(22):10250-5
20479255
J Biol Chem. 1982 Sep 25;257(18):11064-9
6809766
Cancer Res. 2000 Jun 1;60(11):3072-80
10850459
J Biol Chem. 2003 Apr 18;278(16):13875-80
12576469
J Biol Chem. 1998 Dec 18;273(51):34586-93
9852130
Mol Neurobiol. 2001 Aug-Dec;24(1-3):53-64
11831554
FEBS Lett. 1995 Oct 9;373(2):119-22
7589448
J Biol Chem. 2003 Aug 15;278(33):30796-805
12791681
J Biol Chem. 2010 Aug 27;285(35):27360-27371
20573953
Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7031-5
7624364
J Comp Neurol. 2004 Jun 28;474(3):438-52
15174086
J Biol Chem. 2005 Jan 7;280(1):137-45
15504723
J Biol Chem. 2001 Aug 24;276(34):31745-51
11425861
Neuron. 1994 Sep;13(3):583-94
7917292
J Neurosci Res. 2008 Mar;86(4):727-43
17975827
Anal Biochem. 1989 Aug 1;180(2):195-204
2510544
J Biol Chem. 2000 Jun 16;275(24):18594-601
10766765
Genomics. 1997 Jul 1;43(1):43-51
9226371
Nature. 1995 Feb 23;373(6516):715-8
7854457
EMBO J. 1996 Dec 16;15(24):6943-50
9003770
J Biol Chem. 1997 Jul 4;272(27):16884-8
9201996
Proc Natl Acad Sci U S A. 2009 Jul 21;106(29):11995-2000
19587240
Nat Rev Neurosci. 2008 Jan;9(1):26-35
18059411
Mol Cell Neurosci. 1996 Aug;8(2/3):99-111
8954625
J Biol Chem. 1995 Jul 21;270(29):17171-9
7615513
Structure. 2003 Jun;11(6):691-701
12791257
Brain Res Brain Res Rev. 2001 Oct;36(2-3):175-84
11690614
J Biol Chem. 2007 Oct 19;282(42):30346-56
17699524
J Biol Chem. 2005 Dec 30;280(52):42971-7
16267048
Am J Pathol. 1988 Nov;133(2):227-40
2461089