RAPID COMMUNICATION: The heparin-binding 10 kDa fragment of connective tissue growth factor (CTGF) containing module 4 alone stimulates cell adhesion

DK Ball, AW Rachfal, SA Kemper… - Journal of …, 2003 - joe.bioscientifica.com
DK Ball, AW Rachfal, SA Kemper, DR Brigstock
Journal of endocrinology, 2003joe.bioscientifica.com
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 Glu247, was of Mr 10000, was immunoreactive with anti-CTGF [247–
260], bound strongly to heparin, and promoted dose-dependent adhesion of fibroblasts …
Abstract
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 Glu247, was of Mr 10000, 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 Glu247 also promoted cell adhesion. CTGF-mediated cell adhesion was abolished by heparin or EDTA. These data demonstrate the presence of heparinbinding 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.
joe.bioscientifica.com