Isolation of mitogenically active C-terminal truncated pentapeptide of osteogenic growth peptide from human plasma and culture medium of murine osteoblastic cells.
The osteogenic growth peptide (OGP) is a 14-amino acid stromal cell mitogen that stimulates in vivo osteogenesis and hematopoiesis. In the blood circulation and cell culture conditioned medium immunoreactive OGP (irOGP), identified using antibodies raised against the OGP C-terminal region, presents free and bound forms. The bound form consists entirely of the full length peptide. The present study was designed to investigate the identity of free irOGP under nondenaturing conditions. Fresh human serum and culture medium conditioned with murine osteoblastic MC3T3 E1 cells were fractionated using ultrafiltration (3000 molecular weight cut-off). Hydrophobic chromatography of the ultrafiltrate, immunoscreening of chromatographic fractions with antibodies directed against the OGP C-terminal region and amino acid sequencing of immunoreactive peaks demonstrated the presence of two mitogens, the full length OGP and a C-terminal truncated form, OGP(10-14). The OGP(10-14) derived from both serum and conditioned medium, as well as the synthetic pentapeptide [sOGP(10-14)], shared the in vitro OGP proliferative activity. However, in a competitive binding assay, devised to assess the OGP-OGP binding protein (OGPBP) complex formation, sOGP(10-14) failed to compete out radiolabeled OGP from the complex. It is concluded that OGP(10-14) is a naturally occurring human and murine mitogen. In addition, the data suggests that the OGP(10-14) is generated from OGP by proteolytic cleavage upon dissociation of the OGP-OGPBP complexes.
Bab
I
I
Bone Laboratory, Institute of Dental Sciences, Faculty of Dental Medicine, The Hebrew University of Jerusalem, Israel. babi@cc.huji.ac.il
Gavish
H
H
Namdar-Attar
M
M
Muhlrad
A
A
Greenberg
Z
Z
Chen
Y
Y
Mansur
N
N
Shteyer
A
A
Chorev
M
M
D000818:Animals; D001667:Binding, Competitive; D002455:Cell Division; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D017077:Culture Media, Conditioned / Q000737:chemistry; D006133:Growth Substances / Q000097:blood / Q000737:chemistry*; D006657:Histones; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007457:Iodine Radioisotopes; D051379:Mice; D010006:Osteoblasts; D010446:Peptide Fragments / Q000097:blood / Q000302:isolation & purification*; D010455:Peptides / Q000097:blood / Q000737:chemistry*; D011485:Protein Binding; D017421:Sequence Analysis
D017077:Culture Media, Conditioned; D006133:Growth Substances; D006657:Histones; D036341:Intercellular Signaling Peptides and Proteins; D007457:Iodine Radioisotopes; D010446:Peptide Fragments; D010455:Peptides; C074533:osteogenic growth peptide