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ScFv antibody-induced translocation of cell-surface heparan sulfate proteoglycan to endocytic vesicles: Evidence for heparan sulfate epitope specificity and role of both syndecan and glypican.

Wittrup, Anders LU ; Zhang, Sihe LU ; Ten Dam, Gerdy B; van Kuppevelt, Toin H; Bengtson, Per LU ; Johansson, Maria C LU ; Welch, Johanna LU ; Mörgelin, Matthias LU and Belting, Mattias LU (2009) In Journal of Biological Chemistry 284(47). p.32959-32967
Abstract
Cellular uptake of several viruses and polybasic macromolecules requires the expression of cell-surface heparan sulfate proteoglycan (HSPG) through as yet ill-defined mechanisms. We unexpectedly found that among several cell-surface binding scFv anti-HS antibody (alphaHS) clones only one, AO4B08, efficiently translocated macromolecular cargo to intracellular vesicles through induction of HSPG endocytosis. Interestingly, AO4B08-induced PG internalization was strictly dependent on HS 2-O-sulfation and appeared independent on intact N-sulfation. AO4B08 and HIV-tat, i.e. a well-known cell penetrating peptide, were shown to compete for the internalizing PG population. To obtain a more detailed characterization of this pathway, we have developed... (More)
Cellular uptake of several viruses and polybasic macromolecules requires the expression of cell-surface heparan sulfate proteoglycan (HSPG) through as yet ill-defined mechanisms. We unexpectedly found that among several cell-surface binding scFv anti-HS antibody (alphaHS) clones only one, AO4B08, efficiently translocated macromolecular cargo to intracellular vesicles through induction of HSPG endocytosis. Interestingly, AO4B08-induced PG internalization was strictly dependent on HS 2-O-sulfation and appeared independent on intact N-sulfation. AO4B08 and HIV-tat, i.e. a well-known cell penetrating peptide, were shown to compete for the internalizing PG population. To obtain a more detailed characterization of this pathway, we have developed a procedure for the isolation of endocytic vesicles by conjugating AO4B08 with superparamagnetic nano-particles. [35S]sulfate-labelled HSPG was found to accumulate in isolated, AO4B08-containing vesicles, providing first biochemical evidence for intact HSPG co-internalization with its ligand. Further analysis revealed the existence of both syndecan, i.e. a transmembrane HSPG, and glycosylphosphatidyl- inositol anchored glypican in purified vesicles. Importantly, internalized syndecan and glypican were found to colocalize in AO4B08-containing vesicles. Our data establish HSPGs as true internalizing receptors of macromolecular cargo, and indicate that the sorting of cell-surface HSPG to endocytic vesicles is determined by a specific HS epitope that can be carried by both syndecan and glypican core protein. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
284
issue
47
pages
32959 - 32967
publisher
ASBMB
external identifiers
  • wos:000272028400079
  • pmid:19783663
  • scopus:70450225343
ISSN
1083-351X
DOI
10.1074/jbc.M109.036129
language
English
LU publication?
yes
id
035e62bf-9c44-440d-bd07-0397d9c91fa6 (old id 1483121)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/19783663?dopt=Abstract
date added to LUP
2009-10-09 16:09:35
date last changed
2017-11-12 03:19:24
@article{035e62bf-9c44-440d-bd07-0397d9c91fa6,
  abstract     = {Cellular uptake of several viruses and polybasic macromolecules requires the expression of cell-surface heparan sulfate proteoglycan (HSPG) through as yet ill-defined mechanisms. We unexpectedly found that among several cell-surface binding scFv anti-HS antibody (alphaHS) clones only one, AO4B08, efficiently translocated macromolecular cargo to intracellular vesicles through induction of HSPG endocytosis. Interestingly, AO4B08-induced PG internalization was strictly dependent on HS 2-O-sulfation and appeared independent on intact N-sulfation. AO4B08 and HIV-tat, i.e. a well-known cell penetrating peptide, were shown to compete for the internalizing PG population. To obtain a more detailed characterization of this pathway, we have developed a procedure for the isolation of endocytic vesicles by conjugating AO4B08 with superparamagnetic nano-particles. [35S]sulfate-labelled HSPG was found to accumulate in isolated, AO4B08-containing vesicles, providing first biochemical evidence for intact HSPG co-internalization with its ligand. Further analysis revealed the existence of both syndecan, i.e. a transmembrane HSPG, and glycosylphosphatidyl- inositol anchored glypican in purified vesicles. Importantly, internalized syndecan and glypican were found to colocalize in AO4B08-containing vesicles. Our data establish HSPGs as true internalizing receptors of macromolecular cargo, and indicate that the sorting of cell-surface HSPG to endocytic vesicles is determined by a specific HS epitope that can be carried by both syndecan and glypican core protein.},
  author       = {Wittrup, Anders and Zhang, Sihe and Ten Dam, Gerdy B and van Kuppevelt, Toin H and Bengtson, Per and Johansson, Maria C and Welch, Johanna and Mörgelin, Matthias and Belting, Mattias},
  issn         = {1083-351X},
  language     = {eng},
  number       = {47},
  pages        = {32959--32967},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {ScFv antibody-induced translocation of cell-surface heparan sulfate proteoglycan to endocytic vesicles: Evidence for heparan sulfate epitope specificity and role of both syndecan and glypican.},
  url          = {http://dx.doi.org/10.1074/jbc.M109.036129},
  volume       = {284},
  year         = {2009},
}