Mice deficient in N-acetylgalactosamine 4-sulfate 6-o-sulfotransferase are unable to synthesize chondroitin/dermatan sulfate containing N-acetylgalactosamine 4,6-bissulfate residues and exhibit decreased protease activity in bone marrow-derived mast cells.
Chondroitin sulfate (CS) and dermatan sulfate (DS) containing N-acetylgalactosamine 4,6-bissulfate (GalNAc(4,6-SO(4))) show various physiological activities through interacting with numerous functional proteins. N-Acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S-6ST) transfers sulfate from 3'-phosphoadenosine 5'-phosphosulfate to position 6 of N-acetylgalactosamine 4-sulfate in CS or DS to yield GalNAc(4,6-SO(4)) residues. We here report generation of transgenic mice that lack GalNAc4S-6ST. GalNAc4S-6ST-null mice were born normally and fertile. In GalNAc4S-6ST-null mice, GalNAc(4,6-SO(4)) residues in CS and DS disappeared completely, indicating that GalNAc4S-6ST should be a sole enzyme responsible for the synthesis of GalNAc(4,6-SO(4)) residues in both CS and DS. IdoA-GalNAc(4,6-SO(4)) units that account for approximately 40% of total disaccharide units of DS in the liver of the wild-type mice disappeared in the liver DS of GalNAc4S-6ST-null mice without reduction of IdoA content. Bone marrow-derived mast cells (BMMCs) derived from GalNAc4S-6ST-null mice contained CS without GlcA-GalNAc(4,6-SO(4)) units. Tryptase and carboxypeptidase A activities of BMMCs derived from GalNAc4S-6ST-null mice were lower than those activities of BMMCs derived from wild-type mice, although mRNA expression of these mast cell proteases was not altered. Disaccharide compositions of heparan sulfate/heparin contained in the mast cells derived from BMMCs in the presence of stem cell factor were much different from those of heparan sulfate/heparin in BMMCs but did not differ significantly between wild-type mice and GalNAc4S-6ST-null mice. These observations suggest that CS containing GalNAc(4,6-SO(4)) residues in BMMCs may contribute to retain the active proteases in the granules of BMMCs but not for the maturation of BMMCs into connective tissue-type mast cells.
Ohtake-Niimi
Shiori
S
Department of Chemistry, Aichi University of Education, Igaya-cho, Kariya, Aichi 448-8542, Japan.
Kondo
Sachiko
S
Ito
Tatsuro
T
Kakehi
Saori
S
Ohta
Tadayuki
T
Habuchi
Hiroko
H
Kimata
Koji
K
Habuchi
Osami
O
D000116:Acetylgalactosamine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D000818:Animals; D001853:Bone Marrow / Q000201:enzymology* / Q000648:ultrastructure; D002809:Chondroitin Sulfates / Q000096:biosynthesis* / Q000737:chemistry; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D003871:Dermatan Sulfate / Q000096:biosynthesis* / Q000737:chemistry; D004187:Disaccharides / Q000032:analysis; D005091:Exons / Q000235:genetics; D005822:Genetic Vectors; D006025:Glycosaminoglycans / Q000096:biosynthesis*; D008407:Mast Cells / Q000201:enzymology / Q000648:ultrastructure; D051379:Mice; D008854:Microscopy, Electron; D010447:Peptide Hydrolases / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics*; D013154:Spleen / Q000201:enzymology; D015238:Sulfotransferases / Q000172:deficiency* / Q000235:genetics
D017931:DNA Primers; D004187:Disaccharides; D006025:Glycosaminoglycans; D012333:RNA, Messenger; D003871:Dermatan Sulfate; C046735:N-acetylgalactosamine 4-sulfate; D002809:Chondroitin Sulfates; D004247:DNA; C416670:N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase; D015238:Sulfotransferases; D010447:Peptide Hydrolases; D000116:Acetylgalactosamine