The effect of TGF-beta receptor binding peptides on smooth muscle cells.
TGF-beta1 is a potent regulator of vascular smooth muscle cell (VSMC) proliferation, migration, and extracellular matrix (ECM) synthesis. In this study, we selected two peptides, IM-1 and IM-2, that bind to the TGF-beta type II receptor (TGF-beta RII) using phage display. IM-1 and IM-2 bind to the TGF-beta RII, with a K(d) of 1 microM. Like TGF-beta, IM-1 induced VSMC chemotaxis and PAI-1 mRNA expression, as determined using Boyden chambers and real time quantitative PCR. In contrast, IM-2 had no effect on VSMC chemotaxis or PAI-1 induction. Induction of ECM synthesis, involving proteins such as osteopontin and alpha-smooth muscle actin, was determined by ELISA. Osteopontin expression was inhibited by both peptides, but TGF-beta-induced alpha-smooth muscle actin expression could only be inhibited by IM-1. In conclusion, IM-1 activity on VSMC is agonistic with TGF-beta, except for ECM synthesis, whereas the IM-2 peptide is antagonistic for some examined TGF-beta functions.
Michon
Ingrid N
IN
Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands. i.michon@chem.leidenuniv.nl
Penning
Louis C
LC
Molenaar
Tom J M
TJ
van Berkel
Theo J C
TJ
Biessen
Erik A L
EA
Kuiper
Johan
J
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages / Q000378:metabolism; D002478:Cells, Cultured; D002633:Chemotaxis; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000378:metabolism; D007700:Kinetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D053495:Osteopontin; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D051381:Rats; D018125:Receptors, Transforming Growth Factor beta / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000378:metabolism; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000378:metabolism
D000199:Actins; D019151:Peptide Library; D010455:Peptides; D018125:Receptors, Transforming Growth Factor beta; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; C506897:Spp1 protein, rat; D016212:Transforming Growth Factor beta; D053495:Osteopontin