Advanced

Structural basis of glycosaminoglycan modification and of heterotypic interactions of perlecan domain V

Friedrich, M V; Gohring, W; Mörgelin, Matthias LU ; Brancaccio, A; David, G and Timpl, R (1999) In Journal of Molecular Biology 294(1). p.259-270
Abstract
The C-terminal perlecan domain V of about 90 kDa consists of laminin-type G domain modules (LG) (25 kDa) and epidermal growth factor-like modules (EG) (4 kDa) in the tandem arrangement LG1-EG1-EG2-LG2-EG3-EG4-LG3. Several shorter fragments have been prepared by recombinant production in mammalian cells and used to map the single glycosaminoglycan (GAG) substitution site and the binding of several carbohydrate and protein ligands. This identified a Ser3511 residue located in a short link region between EG4 and LG3 as being involved in GAG attachment. Electron microscopy provided evidence that the same substitution exists in tissue forms of perlecan. Heparan sulphate attached to this site was shown to bind to the alpha1LG4 module of... (More)
The C-terminal perlecan domain V of about 90 kDa consists of laminin-type G domain modules (LG) (25 kDa) and epidermal growth factor-like modules (EG) (4 kDa) in the tandem arrangement LG1-EG1-EG2-LG2-EG3-EG4-LG3. Several shorter fragments have been prepared by recombinant production in mammalian cells and used to map the single glycosaminoglycan (GAG) substitution site and the binding of several carbohydrate and protein ligands. This identified a Ser3511 residue located in a short link region between EG4 and LG3 as being involved in GAG attachment. Electron microscopy provided evidence that the same substitution exists in tissue forms of perlecan. Heparan sulphate attached to this site was shown to bind to the alpha1LG4 module of laminin-1, indicating a role in basement membrane assembly and cell-matrix interactions. This site is also close to an Asn-Asp bond which is readily cleaved by an endogenous protease that depends on the presence of Asp and the LG2 module. A weak heparin binding site was shown to include the EG2 module, which contains five basic residues. Binding to sulphatides and the alpha-dystroglycan receptor was much stronger and required at least two LG modules. However, single LG modules appear to be sufficient for the interaction with the laminin-nidogen complex, while EG3-4 and some flanking regions are apparently involved in fibulin-2 binding. These observations indicate that a complex modular structure is required for domain V in order to provide a rich repertoire of potential biological functions. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
basement membranes, greek small letter alpha-dystroglycan, carbohydrate ligands, recombinant production, site-directed mutagenesis
in
Journal of Molecular Biology
volume
294
issue
1
pages
259 - 270
publisher
Elsevier
external identifiers
  • pmid:10556044
  • scopus:0345148341
ISSN
1089-8638
DOI
10.1006/jmbi.1999.3259
language
English
LU publication?
yes
id
79f4f452-65e8-44bf-9f8a-6ffbe7cddc78 (old id 1115461)
date added to LUP
2008-07-08 09:47:21
date last changed
2017-01-01 06:42:07
@article{79f4f452-65e8-44bf-9f8a-6ffbe7cddc78,
  abstract     = {The C-terminal perlecan domain V of about 90 kDa consists of laminin-type G domain modules (LG) (25 kDa) and epidermal growth factor-like modules (EG) (4 kDa) in the tandem arrangement LG1-EG1-EG2-LG2-EG3-EG4-LG3. Several shorter fragments have been prepared by recombinant production in mammalian cells and used to map the single glycosaminoglycan (GAG) substitution site and the binding of several carbohydrate and protein ligands. This identified a Ser3511 residue located in a short link region between EG4 and LG3 as being involved in GAG attachment. Electron microscopy provided evidence that the same substitution exists in tissue forms of perlecan. Heparan sulphate attached to this site was shown to bind to the alpha1LG4 module of laminin-1, indicating a role in basement membrane assembly and cell-matrix interactions. This site is also close to an Asn-Asp bond which is readily cleaved by an endogenous protease that depends on the presence of Asp and the LG2 module. A weak heparin binding site was shown to include the EG2 module, which contains five basic residues. Binding to sulphatides and the alpha-dystroglycan receptor was much stronger and required at least two LG modules. However, single LG modules appear to be sufficient for the interaction with the laminin-nidogen complex, while EG3-4 and some flanking regions are apparently involved in fibulin-2 binding. These observations indicate that a complex modular structure is required for domain V in order to provide a rich repertoire of potential biological functions.},
  author       = {Friedrich, M V and Gohring, W and Mörgelin, Matthias and Brancaccio, A and David, G and Timpl, R},
  issn         = {1089-8638},
  keyword      = {basement membranes,greek small letter alpha-dystroglycan,carbohydrate ligands,recombinant production,site-directed mutagenesis},
  language     = {eng},
  number       = {1},
  pages        = {259--270},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Biology},
  title        = {Structural basis of glycosaminoglycan modification and of heterotypic interactions of perlecan domain V},
  url          = {http://dx.doi.org/10.1006/jmbi.1999.3259},
  volume       = {294},
  year         = {1999},
}