Enzymatically active N-deacetylase/N-sulfotransferase-2 is present in liver but does not contribute to heparan sulfate N-sulfation.
Heparan sulfate (HS) proteoglycans influence embryonic development through interactions with growth factors and morphogens. The interactions depend on HS structure, which is largely determined during biosynthesis by Golgi enzymes. NDST (glucosaminyl N-deacetylase/N-sulfotransferase), responsible for HS N-sulfation, is a key enzyme directing further modifications including O-sulfation. To elucidate the roles of the different NDST isoforms in HS biosynthesis, we took advantage of mice with targeted mutations in NDST1 and NDST2 and used liver as our model organ. Of the four NDST isoforms, only NDST1 and NDST2 transcripts were shown to be expressed in control liver. The absence of NDST1 or NDST2 in the knock-out mice did not affect transcript levels of other NDST isoforms or other HS modification enzymes. Although the sulfation level of HS synthesized in NDST1-/- mice was drastically lowered, liver HS from wild-type mice, from NDST1+/-, NDST2-/-, and NDST1+/- / NDST2-/- mice all had the same structure despite greatly reduced NDST enzyme activity (30% of control levels in NDST1+/- / NDST2-/- embryonic day 18.5 embryos). Enzymatically active NDST2 was shown to be present in similar amounts in wild-type, NDST1-/-, and NDST1+/- embryonic day 18.5 liver. Despite the substantial contribution of NDST2 to total NDST enzyme activity in embryonic day 18.5 liver (approximately 40%), its presence did not appear to affect HS structure as long as NDST1 was also present. In NDST1-/- embryonic day 18.5 liver, in contrast, NDST2 was responsible for N-sulfation of the low sulfated HS. A tentative model to explain these results is presented.
Ledin
Johan
J
Department of Medical Biochemistry and Microbiology, Uppsala University, SE-751 23 Uppsala, Sweden.
Ringvall
Maria
M
Thuveson
Maria
M
Eriksson
Inger
I
Wilén
Maria
M
Kusche-Gullberg
Marion
M
Forsberg
Erik
E
Kjellén
Lena
L
D000581:Amidohydrolases / Q000096:biosynthesis*; D000818:Animals; D018507:Gene Expression Regulation, Developmental; D005838:Genotype; D006025:Glycosaminoglycans / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry* / Q000378:metabolism; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009154:Mutation; D020033:Protein Isoforms; D015238:Sulfotransferases / Q000096:biosynthesis*; D013997:Time Factors
D000581:Amidohydrolases; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D020033:Protein Isoforms; D015238:Sulfotransferases; C491855:Ndst2 protein, mouse; C021350:heparitin sulfotransferase