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147 Copyright c~ 1987 by Academic Press, Inc. All rights of reproduction in any form reserved. 22 PLATELET-DERIVED GROWTH FACTOR  the estimates may differ from the actual concentration if the Kd of the competing molecule for the receptor is significantly higher or lower than 1251-labeled PDGF. Acknowledgment The author is thankful to Rose Erler for preparation of the manuscript.  S t r u c t u r a l a n d F u n c t i o n a l I d e n t i f i c a t i o n o f Platelet-Derived Growth Factor-Like Proteins Produced by Mammalian C el l s B y HARRY N .
Natl. Acad. Sci. A. 78, 2918 (1981). E. P. , Proc. Natl. Acad. Sci. A. 78, 3373 (1981). 6 S. G. , Proc. Natl. Acad. Sci. A. 80, 731 (1983). M E T H O D S IN E N Z Y M O L O G Y , V O L . 147 Copyright c~ 1987 by Academic Press, Inc. All rights of reproduction in any form reserved.  IDENTIFICATION OF P D G F - L I K E PROTEINS 23 biologically active PDGF. This important question has been resolved recently by the demonstration that, in SSV-transformed cells, p28 ~'-' undergoes a series of discrete processing steps including dimer formation and proteolytic cleavage to yield molecules structurally and immunologically resembling the disulfide-linked dimeric forms of PDGF5 More recent studies have shown that this processed product is secreted by SSV-transformed fibroblasts into their culture media in a biologically active form which is recognized by PDGF antisera, s These studies established conclusively that the biologically active SSV one gene product is a homodimer, consisting of two PDGF-2 chains linked together by disulfide bonds.
Scher, W. J. Pledger, P. Martin, H. N. Antoniades, and C. D. Stiles, J. Cell. Physiol. 97, 371 (1978). 17 W. H. Kirsten and L. A. Myer, J. Natl. Cancerlnst. ) 39, 311 (1967). is L. J. Gudas, J. P. Singh, and C. D. Stiles, Cold Spring Harbor ConJ] Cell Proliferation 10, 229 (1983). ~9 H. N. A n t o n i a d e s , C. D. Scher, and C. D. Stiles, Proc. Natl. Acad. Sci. A. 76, 1809 (1979). 16 PLATELET-DERIVED GROWTH FACTOR  Outdated Platelets I I Platelet Pellet I I 1M NaCl Platelet Extract I I Dialysis I I CM-Sephadex I I BioGeI-P-150 I I Reversed-Phase HP LC / / / HPLC Molecular Sizing Chromatography I I PDGF \ \ \ lodination I I I HPLC Molecular Sizing Chromatography I I 1251-PDGF FIG.