The heparin-binding 10 kDa fragment of connective tissue growth factor (CTGF) containing module 4 alone stimulates cell adhesion.
Connective tissue growth factor (CTGF) is a 349-residue mosaic protein that contains four structural modules implicated in protein-protein interactions. To address the functionality of residues 247-349 (containing module 4 alone), this region of CTGF was produced as a maltose binding protein (MBP) fusion protein in E. coli. After removal of MBP, recombinant CTGF commenced at Glu(247), was of M(r) 10 000, was immunoreactive with anti-CTGF[247-260], bound strongly to heparin, and promoted dose-dependent adhesion of fibroblasts, myofibroblasts, endothelial cells, and epithelial cells. An 8 kDa presumptive C-terminally truncated form of CTGF commencing at Glu(247) also promoted cell adhesion. CTGF-mediated cell adhesion was abolished by heparin or EDTA. These data demonstrate the presence of heparin-binding and cell-adhesion motifs within the C-terminal 103 residues of CTGF and show that CTGF-mediated cell adhesion is heparin-and divalent cation-dependent. Thus, CTGF isoforms comprising essentially module 4 are intrinsically functional in the absence of the other constituent modules of CTGF.
Ball
D K
DK
Department of Surgery, The Ohio State University, Columbus, Ohio 43212, USA.
Rachfal
A W
AW
Kemper
S A
SA
Brigstock
D R
DR
D016475:3T3 Cells / Q000166:cytology / Q000187:drug effects; D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D003239:Connective Tissue Cells / Q000166:cytology* / Q000187:drug effects; D055513:Connective Tissue Growth Factor; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology; D004727:Endothelium / Q000166:cytology / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism / Q000494:pharmacology*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism / Q000494:pharmacology*; D008099:Liver / Q000166:cytology / Q000187:drug effects; D051379:Mice; D010446:Peptide Fragments / Q000494:pharmacology; D011485:Protein Binding; D011994:Recombinant Proteins / Q000494:pharmacology; D013268:Stimulation, Chemical
D000925:Anticoagulants; C526317:CCN2 protein, human; C526318:CCN2 protein, mouse; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D055513:Connective Tissue Growth Factor; D006493:Heparin; D004492:Edetic Acid