title: Splicing determines the glycosylation state of ameloblastin.
abstract: In developing porcine enamel, the space between enamel rods selectively binds lectins and ameloblastin (Ambn) N-terminal antibodies. We tested the hypothesis that ameloblastin N-terminal cleavage products are glycosylated. Assorted Ambn cleavage products showed positive lectin staining by peanut agglutinin (PNA), Maclura pomifera agglutinin (MPA), and Limulus polyphemus agglutinin (LPA), suggesting the presence of an O-linked glycosylation containing galactose (Gal), N-acetylgalactosamine (GalNAc), and sialic acid. Edman sequencing of the lectin-positive bands gave the Ambn N-terminal sequence: VPAFPRQPGTXGVASLXLE. The blank cycles for Pro(11) and Ser(17) confirmed that these residues are hydroxylated and phosphorylated, respectively. The O-glycosylation site was determined by Edman sequencing of pronase-digested Ambn, which gave HPPPLPXQPS, indicating that Ser(86) is the site of the O-linked glycosylation. This modification is within the 15-amino-acid segment (73-YEYSLPVHPPPLPSQ-87) deleted by splicing in the mRNA encoding the 380-amino-acid Ambn isoform. We conclude that only the N-terminal Ambn products derived from the 395-Ambn isoform are glycosylated.
authors: Department of Biologic and Materials Sciences, Dental Research Lab, 1210 Eisenhower Place, Ann Arbor, MI 48108, USA.
authors: K
authors: K
authors: Kobayashi
authors: Y
authors: Y
authors: Yamakoshi
authors: J C-C
authors: JC
authors: Hu
authors: K
authors: K
authors: Gomi
authors: T
authors: T
authors: Arai
authors: M
authors: M
authors: Fukae
authors: P H
authors: PH
authors: Krebsbach
authors: J P
authors: JP
authors: Simmer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003746:Dental Enamel Proteins / Q000235:genetics* / Q000378:metabolism*; D006031:Glycosylation; D037102:Lectins / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D020033:Protein Isoforms; D011499:Protein Processing, Post-Translational; D012326:RNA Splicing; D013552:Swine
chemical_list: D003746:Dental Enamel Proteins; D037102:Lectins; D010446:Peptide Fragments; D020033:Protein Isoforms; C490891:ameloblastin protein, Sus scrofa