Rapid chromatographic isolation and immunoblot characterization of immunoreactive fibroblast growth factor-related polypeptides from various tissues.
Procedures to rapidly isolate fibroblast growth factor (FGF)-like activity from a number of tissue sources (lung, plasma, brain, ovary, corpus luteum, pituitary, chondrosarcoma) of bovine, porcine or rat origin are described. In addition, immunoblotting experiments using well characterized and specific rabbit polyclonal anti-fibroblast growth factor beta (anti-FGF-beta) sera have been performed. Besides documenting the first report of the isolation of FGF-beta from bovine lung and plasma, these studies provide evidence for the existence of higher-molecular-mass proteins with FGF-beta-like immunoreactivity. For example, in addition to new truncated forms of the acidic and basic FGF (FGF-alpha and FGF-beta), respectively, other higher-molecular-mass immunoreactive proteins were detected in bovine, pig and rat brain, and in rat chondrosarcoma. The tissue distribution of these immunoreactive proteins and their competitive inhibition characteristics mitigate against the possibility that the polyclonal antisera are cross-reacting non-specifically with common cellular proteins. Rather, the data suggest that the immunoblotting technique is either detecting other proteins structurally related to FGF-beta or alternatively FGF-beta is strongly bound to specific carrier proteins (e.g. heparan sulphate proteoglycan fragments) associated with their transport and recognition at the cellular level.
Bertolini
J
J
Department of Biochemistry, Monash University, Clayton, Victoria Australia.
Guthridge
M
M
Hearn
M T
MT
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005346:Fibroblast Growth Factors / Q000302:isolation & purification*; D010455:Peptides / Q000302:isolation & purification*; D051381:Rats; D013552:Swine
D010455:Peptides; D005346:Fibroblast Growth Factors