Premium
3‐O‐sulfation provides high affinity binding to neuropilin‐1 and modulates endothelial cell sprouting and neuronal growth cone collapse (607.7)
Author(s) -
Thacker Bryan,
Lawrence Roger,
Seamen Emylie,
Liu JIan,
Esko Jeffrey
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.607.7
Subject(s) - sulfation , heparan sulfate , chemistry , biochemistry , neuropilin 1 , sulfotransferase , semaphorin , perlecan , sulfate , glycosaminoglycan , receptor , biology , vascular endothelial growth factor , organic chemistry , cancer research , vegf receptors
Heparan sulfate structural heterogeneity is driven, in part, by the placement of sulfate groups at various positions in the polysaccharide. While many ligands bind to heparan sulfate without strict requirements for the positions of sulfate groups, binding of a small number of known ligands is influenced by the presence of a sulfate at the C3 position of a glucosamine residue. In mammals, seven enzymes can catalyze the addition of 3‐O‐sulfate groups, suggesting the possibility of other, previously unidentified, ligands whose binding is influenced by 3‐O‐sulfated sequences. To identify these ligands, affinity matrices were created using CHO‐S heparan sulfate, with and without modification by recombinant 3‐O‐sulfotransferases. Serum was fractionated on the matrices and several proteins binding specifically to the 3‐O‐sulfated resins were identified by mass spectrometry. Neuropilin‐1, a modulator of angiogenesis and axonal guidance, bound specifically to affinity matrices with 3‐O‐sulfation. In addition, high affinity binding of neuropilin‐1 to the cell surface required 3‐O‐sulfation. In cell culture, 3‐O‐sulfation modulated two neuropilin‐1 dependent processes. (1) In VEGF‐induced endothelial cell sprouting, 3‐O‐sulfation endowed CHO‐S heparan sulfate with greater inhibitory potential than native heparan sulfate. (2) In semaphorin‐3a‐induced neuronal growth cone collapse, a 3‐O‐sulfated heparan sulfate dodecamer was a potent inhibitor of collapse. An identical dodecamer lacking 3‐O‐sulfation inhibited with much less efficiency. Furthermore, genetic removal of a 3‐O‐sulfotransferase (Hs3st‐2) attenuated the response of the growth cone to semaphorin‐3a. In conclusion, these results demonstrate that 3‐O‐sulfation endows heparan sulfate with enhanced affinity to neuropilin‐1 and modulates two neuropilin‐1 dependent processes, angiogenesis and axonal guidance. Grant Funding Source : Supported by GM93131 and T32CA067754