The C-terminal peptide of thrombospondin-4 stimulates erythroid cell proliferation.
Erythropoietin (EPO) stimulates the production of small erythroid cell stimulating factors (molecular weight <5 kDa) in cultures of bone marrow endothelial cells. We identified a fragment of thrombospondin-4 (TSP-4) as an EPO-stimulated protein in endothelial cell lysates. Pre-incubation of the low molecular weight fractions from supernatants of EPO-treated umbilical cord endothelial cells (HUVEC) with antibodies against the C-terminal residues of TSP-1,2 and TSP-4 decreased the erythroid cell stimulating activity. The C-terminal TSP-1 section corresponding to a molecular weight lower than 6 kDa has the integrin-associated protein binding motif VVM. The corresponding TSP-4 fragment, lacking the three residue sequence VVM, has a distinctive acidic peptide comprising the last 21 amino acids (C21) with the characteristics of an amphipathic helix. C21 stimulated thymidine incorporation into bovine erythroid cells, increased cell numbers in cultures of cord blood CD36+ erythroid precursors and skin fibroblasts, and decreased HUVEC proliferation. SC21, a homologous peptide of identical amino acid composition but with interchanged residues, was non-amphipathic and had no erythroid cell stimulating activity.
Endocrine Laboratory, McGill University Health Centre, Montreal, Canada. luis.f.congote@muhc.mcgill.ca
Luis F
LF
Congote
Marcos R
MR
Difalco
Bernard F
BF
Gibbs
D000595:Amino Acid Sequence; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002417:Cattle; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D002942:Circular Dichroism; D004305:Dose-Response Relationship, Drug; D004306:Dose-Response Relationship, Immunologic; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D041905:Erythroid Cells / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D017434:Protein Structure, Tertiary; D019700:Thrombospondin 1 / Q000378:metabolism; D019699:Thrombospondins / Q000737:chemistry* / Q000378:metabolism; D013936:Thymidine / Q000737:chemistry; D013997:Time Factors; D000099262:Thrombospondin 2
D007797:Laminin; D010455:Peptides; D019700:Thrombospondin 1; D019699:Thrombospondins; D013936:Thymidine; D000099262:Thrombospondin 2; C101864:thrombospondin 4