Heparan sulfate composition of alternatively spliced CD44 fusion proteins.
Prior analyses of recombinant CD44 fusion proteins have indicated that combinatorial splicing of variant exons exerts distal effects on chondroitin sulfate content and structure, which may regulate the biological properties of the respective CD44 isoforms. The consequences of splicing of variant exons V4-7 on the heparan sulfate moieties were therefore examined, utilizing recombinant chimeras containing exons V3 and V8-10, engineered with or without exons V4-7 and expressed as Ig fusion proteins in COS cells. Splicing of exons V4-7, though they contain no consensus motifs for glycosaminoglycan assembly, resulted in markedly increased polymer sulfation levels of the heparan sulfates. The sulfate groups of both the CD44 V3-10 and V3,8-10 isoforms occurred as di- and tri-sulfated dissacharide units and were restricted to one N-sulfated block domain within the polymers. Compared to native human keratinocyte CD44, the recombinant heparan sulfates were relatively low in sulfate content. Our data indicate that variant exon V4-7 splicing exerts distal effects on the composition of this glycosaminoglycan. These effects may regulate those functions that are mediated through the heparan sulfate moieties, such as the binding of growth factors.
Piepkorn
M
M
Departments of Medicine (Dermatology) and Pathology, University of Washington School of Medicine, Seattle, Washington, USA. mpiepkor@u.washington.edu
Hovingh
P
P
Bennett
K L
KL
Linker
A
A
D017398:Alternative Splicing / Q000235:genetics*; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000032:analysis; D002852:Chromatography, Ion Exchange; D004187:Disaccharides / Q000032:analysis / Q000378:metabolism; D005091:Exons / Q000235:genetics; D006497:Heparan Sulfate / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007231:Infant, Newborn; D015603:Keratinocytes; D009844:Oligosaccharides / Q000032:analysis / Q000378:metabolism; D011108:Polymers / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000378:metabolism; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D017384:Sequence Deletion
D002809:Chondroitin Sulfates; D004187:Disaccharides; D006497:Heparan Sulfate; D018960:Hyaluronan Receptors; D009844:Oligosaccharides; D011108:Polymers; D011133:Polysaccharide-Lyases; D020033:Protein Isoforms; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; C021349:heparitinsulfate lyase