Apoptotic cell thrombospondin-1 and heparin-binding domain lead to dendritic-cell phagocytic and tolerizing states

Blood ◽  
2006 ◽  
Vol 108 (10) ◽  
pp. 3580-3589 ◽  
Author(s):  
Alon Krispin ◽  
Yaniv Bledi ◽  
Mizhir Atallah ◽  
Uriel Trahtemberg ◽  
Inna Verbovetski ◽  
...  
Keyword(s):  
Dendritic Cell ◽  
Apoptotic Cell ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Binding Domain ◽  
Apoptotic Cells ◽  
Heparin Binding ◽  
Proteomic Approach ◽  
Thrombospondin 1 ◽  
Heparin Binding Domain

Abstract Apoptotic cells were shown to induce dendritic cell immune tolerance. We applied a proteomic approach to identify molecules that are secreted from apoptotic monocytes, and thus may mediate engulfment and immune suppression. Supernatants of monocytes undergoing apoptosis were collected and compared using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and differentially expressed proteins were identified using tandem mass spectrometry. Thrombospondin-1 (TSP-1) and its cleaved 26-kDa heparin-binding domain (HBD) were identified. We show that TSP-1 is expressed upon induction of monocyte apoptosis in a caspase-dependent pattern and the HBD is cleaved by chymotrypsin-like serine protease. We further show that CD29, CD36, CD47, CD51, and CD91 simultaneously participate in engulfment induction and generation of an immature dendritic cell (iDC) tolerogenic and phagocytic state. We conclude that apoptotic cell TSP-1, and notably its HBD, creates a signalosome in iDCs to improve engulfment and to tolerate engulfed material prior to the interaction with apoptotic cells.

scholarly journals Cellular Internalization and Degradation of Thrombospondin-1 Is Mediated by the Amino-terminal Heparin Binding Domain (HBD)

1997 ◽  
Vol 272 (10) ◽  
pp. 6784-6791 ◽  
Author(s):  
Irina Mikhailenko ◽  
Dmitry Krylov ◽  
Kelley McTigue Argraves ◽  
David D. Roberts ◽  
Gene Liau ◽  
...  
Keyword(s):  
Binding Domain ◽  
Cellular Internalization ◽  
Heparin Binding ◽  
Amino Terminal ◽  
Thrombospondin 1 ◽  
Heparin Binding Domain

Thrombospondin-1 binds to polyhistidine with high affinity and specificity

2000 ◽  
Vol 347 (2) ◽  
pp. 469-473 ◽  
Author(s):  
Vijay K. VANGURI ◽  
Shuxia WANG ◽  
Svetlana GODYNA ◽  
Sripriya RANGANATHAN ◽  
Gene LIAU
Keyword(s):  
Solid Phase ◽  
Imidazole Ring ◽  
Binding Domain ◽  
Heparin Binding ◽  
Surface Receptors ◽  
High Affinity ◽  
Thrombospondin 1 ◽  
Heparin Binding Domain ◽  
Solid Phase Binding ◽  
And Migration

Thrombospondin-1 (TSP1) is a secreted trimeric glycoprotein of 450 kDa with demonstrated effects on cell growth, adhesion and migration. Its complex biological activity is attributed to its ability to bind to cell-surface receptors, growth factors and extracellular-matrix proteins. In this study, we used a 125I solid-phase binding assay to demonstrate that TSP1 binds specifically to proteins containing polyhistidine stretches. Based on studies with three different six-histidine-containing recombinant proteins, we derived an average dissociation constant of 5 nM. The binding of 125I-labelled TSP1 to these proteins was inhibited by peptides containing histidine residues, with the degree of competition being a function of the number of histidines within the peptide. Binding was not inhibited by excess histidine or imidazole, indicating that the imidazole ring is not sufficient for recognition by TSP1. Heparin was a potent inhibitor of binding with a Ki of 50 nM, suggesting that the heparin-binding domain of TSP1 may be involved in this interaction. This was confirmed by the ability of a recombinant heparin-binding domain of TSP1 to directly compete for TSP1 binding to polyhistidine-containing proteins. Affinity chromatography with a polyhistidine-containing peptide immobilized on agarose revealed that TSP1 in platelet releasates is the major polypeptide retained on the six-histidine-peptide column. We conclude that TSP1 contains a high-affinity binding site for polyhistidine and this is likely to be the molecular basis for the observed binding of TSP1 to histidine-rich glycoprotein. The possibility that other polyhistidine-containing proteins also interact with TSP1 warrants further study.

A recombinant NH2-terminal heparin-binding domain of the adhesive glycoprotein, thrombospondin-1, promotes endothelial tube formation and cell survival: a possible role for syndecan-4 proteoglycan

2002 ◽  
Vol 21 (4) ◽  
pp. 311-324 ◽  
Author(s):  
Marianna A Ferrari do Outeiro-Bernstein ◽  
Sara Santana Nunes ◽  
Ana Carolina Magalhães Andrade ◽  
Tercia Rodrigues Alves ◽  
Chantal Legrand ◽  
...  
Keyword(s):  
Cell Survival ◽  
Tube Formation ◽  
Binding Domain ◽  
Heparin Binding ◽  
Endothelial Tube Formation ◽  
Thrombospondin 1 ◽  
Heparin Binding Domain

Overlap of IGF- and heparin-binding sites in rat IGF-binding protein-5

2000 ◽  
Vol 24 (1) ◽  
pp. 43-51 ◽  
Author(s):  
H Song ◽  
J Beattie ◽  
IW Campbell ◽  
GJ Allan
Keyword(s):  
Binding Site ◽  
Conformational Changes ◽  
Binding Protein ◽  
Binding Activity ◽  
Binding Domain ◽  
Site Directed Mutagenesis ◽  
Heparin Binding ◽  
Igf I ◽  
Heparin Binding Domain ◽  
Igf Binding

Using site-directed mutagenesis, we have undertaken a study of a potential IGF-binding site in the C-terminal domain of rat IGFBP-5, lying close to or within a previously described heparin-binding domain (residues 201-218) in this protein. After analysis of binding activity using three different methods - ligand blotting, solution phase equilibrium binding and biosensor measurement of real-time on- and off-rates - we report that the mutation of two highly conserved residues within this region (glycine 203 and glutamine 209) reduces the affinity of the binding protein for both IGF-I and IGF-II, while having no effect on heparin binding. In addition, we confirm that mutation of basic residues within the heparin-binding domain (R201L, K202E, K206Q and R214A) results in a protein that has attenuated heparin binding but shows only a small reduction in affinity for IGF-I and -II. Previous findings have described the reduction in affinity of IGFBP-5 for IGFs that occurs after complexation of the binding protein with heparin or other components of the extracellular matrix (ECM) and have postulated that such an interaction may result in conformational changes in protein structure, affecting subsequent IGF interaction. Our data suggesting potential overlap of heparin- and IGF-binding domains argue for a more direct effect of ECM modulation of the affinity of IGFBP-5 for ligand by partial occlusion of the IGF-binding site after interaction with ECM.

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