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.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1483121
- author
- 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
- organization
- publishing date
- 2009
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 284
- issue
- 47
- pages
- 32959 - 32967
- publisher
- American Society for Biochemistry and Molecular Biology
- 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
- 2016-04-01 11:40:52
- date last changed
- 2022-01-26 08:39:19
@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 = {{American Society for Biochemistry and Molecular Biology}},
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}},
doi = {{10.1074/jbc.M109.036129}},
volume = {{284}},
year = {{2009}},
}