Purification and identification of a BMP-like factor from bovine serum.
Myogenic differentiation is suppressed in vitro by unknown factors present in fetal bovine serum (FBS). We found that specific inhibitors of bone morphogenetic proteins (BMPs) stimulated myogenic differentiation even in the presence of 20% FBS, which in turn activated specific BMP signaling. Moreover, these specific BMP inhibitors blocked maturation of osteoblastic cells induced by FBS, indicating that BMP-like factor(s) in serum regulate both myogenic and osteoblastic differentiation. The factor identified had an apparent molecular weight (Mw) of over 100kDa on a Superdex 200 column for molecular sieving HPLC, but an apparent Mw of 33kDa on SDS-PAGE under non-reducing conditions. Analysis of a purified preparation from FBS (5L) by liquid chromatography-tandem mass spectrometry revealed the presence of an amino acid sequence conserved between mature human and murine BMP-4. This is the first study to show that BMP-4 is present in FBS as a large complex.
Kodaira
Kunihiko
K
Division of Pathophysiology, Saitama Medical School, 1397-1 Yamane, Hidaka-shi, Saitama 350-1241, Japan.
Imada
Mana
M
Goto
Masaaki
M
Tomoyasu
Akihiro
A
Fukuda
Toru
T
Kamijo
Ryutaro
R
Suda
Tatsuo
T
Higashio
Kanji
K
Katagiri
Takenobu
T
D000595:Amino Acid Sequence; D000818:Animals; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000097:blood* / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002454:Cell Differentiation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D055427:Growth Differentiation Factor 2; D055412:Growth Differentiation Factors; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D015398:Signal Transduction
C525809:BMP4 protein, human; C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; C525887:GDF2 protein, human; C526108:Gdf2 protein, mouse; D055427:Growth Differentiation Factor 2; D055412:Growth Differentiation Factors