Expression of three recombinant homodimeric isoforms of PDGF in saccharomyces cerevisiae: Evidence for difference in receptor binding and functional activities

Arne Östman; Gudrun Bäckström; Noel Fong; Christer Betsholtz; Christer Wernstedt; Ulf Hellman; Bengt Westermark; Pablo Valenzuela; Carl Henrik Heldin

doi:10.3109/08977198908998003

Expression of three recombinant homodimeric isoforms of PDGF in saccharomyces cerevisiae: Evidence for difference in receptor binding and functional activities

Arne Östman, Gudrun Bäckström, Noel Fong, Christer Betsholtz, Christer Wernstedt, Ulf Hellman, Bengt Westermark, Pablo Valenzuela, Carl Henrik Heldin

Life Sciences School

Research output: Contribution to journal › Article › peer-review

74 Citations (Scopus)

Abstract

Three recombinant homodimeric isoforms of platelet-derived growth factor (PDGF) were produced and purified in milligram quantities by expression of PDGF A-and B-chains in yeast cells. Structural analysis of the purified short and long variants of PDGF-AA (PDGF-AA_S and PDGF-AA_L) and PDGF-BB showed that they had been properly processed and assembled into dimers. PDGF-AA_s and PDGF-AA_L were found to bind only to the PDGF A-type receptor on human fibroblasts, with affinities of 0.1 and 0.2 uM, respectively. PDGF-BB bound to cells with A-and B-type receptors and to cells with B-type receptor only with affinities of 0.6 nM in both cases. Each fibroblast appeared to express about 4-5 times more B-type receptors than A-type receptors. The maximal mitogenic response to PDGF-BB of human fibroblasts was almost 2-fold higher than that induced by either of the two PDGF-AA forms. The three isoforms of PDGF also stimulated growth in soft agar of human fibroblasts with PDGF-BB inducing a higher maximal response.

Original language	English
Pages (from-to)	271-281
Number of pages	11
Journal	Growth Factors
Volume	1
Issue number	3
DOIs	https://doi.org/10.3109/08977198908998003
Publication status	Published - 1 Jan 1989

Keywords

Isoforms
PDGF
Yeast expression

ASJC Scopus subject areas

Endocrinology
Clinical Biochemistry
Cell Biology

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10.3109/08977198908998003

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Östman, A., Bäckström, G., Fong, N., Betsholtz, C., Wernstedt, C., Hellman, U., Westermark, B., Valenzuela, P., & Heldin, C. H. (1989). Expression of three recombinant homodimeric isoforms of PDGF in saccharomyces cerevisiae: Evidence for difference in receptor binding and functional activities. Growth Factors, 1(3), 271-281. https://doi.org/10.3109/08977198908998003

Östman, Arne ; Bäckström, Gudrun ; Fong, Noel ; Betsholtz, Christer ; Wernstedt, Christer ; Hellman, Ulf ; Westermark, Bengt ; Valenzuela, Pablo ; Heldin, Carl Henrik. / Expression of three recombinant homodimeric isoforms of PDGF in saccharomyces cerevisiae : Evidence for difference in receptor binding and functional activities. In: Growth Factors. 1989 ; Vol. 1, No. 3. pp. 271-281.

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abstract = "Three recombinant homodimeric isoforms of platelet-derived growth factor (PDGF) were produced and purified in milligram quantities by expression of PDGF A-and B-chains in yeast cells. Structural analysis of the purified short and long variants of PDGF-AA (PDGF-AAS and PDGF-AAL) and PDGF-BB showed that they had been properly processed and assembled into dimers. PDGF-AAs and PDGF-AAL were found to bind only to the PDGF A-type receptor on human fibroblasts, with affinities of 0.1 and 0.2 uM, respectively. PDGF-BB bound to cells with A-and B-type receptors and to cells with B-type receptor only with affinities of 0.6 nM in both cases. Each fibroblast appeared to express about 4-5 times more B-type receptors than A-type receptors. The maximal mitogenic response to PDGF-BB of human fibroblasts was almost 2-fold higher than that induced by either of the two PDGF-AA forms. The three isoforms of PDGF also stimulated growth in soft agar of human fibroblasts with PDGF-BB inducing a higher maximal response.",

keywords = "Isoforms, PDGF, Yeast expression",

author = "Arne {\"O}stman and Gudrun B{\"a}ckstr{\"o}m and Noel Fong and Christer Betsholtz and Christer Wernstedt and Ulf Hellman and Bengt Westermark and Pablo Valenzuela and Heldin, {Carl Henrik}",

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Östman, A, Bäckström, G, Fong, N, Betsholtz, C, Wernstedt, C, Hellman, U, Westermark, B, Valenzuela, P & Heldin, CH 1989, 'Expression of three recombinant homodimeric isoforms of PDGF in saccharomyces cerevisiae: Evidence for difference in receptor binding and functional activities', Growth Factors, vol. 1, no. 3, pp. 271-281. https://doi.org/10.3109/08977198908998003

Expression of three recombinant homodimeric isoforms of PDGF in saccharomyces cerevisiae : Evidence for difference in receptor binding and functional activities. / Östman, Arne; Bäckström, Gudrun; Fong, Noel; Betsholtz, Christer; Wernstedt, Christer; Hellman, Ulf; Westermark, Bengt; Valenzuela, Pablo; Heldin, Carl Henrik.

In: Growth Factors, Vol. 1, No. 3, 01.01.1989, p. 271-281.

Research output: Contribution to journal › Article › peer-review

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AU - Bäckström, Gudrun

AU - Fong, Noel

AU - Betsholtz, Christer

AU - Wernstedt, Christer

AU - Hellman, Ulf

AU - Westermark, Bengt

AU - Valenzuela, Pablo

AU - Heldin, Carl Henrik

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N2 - Three recombinant homodimeric isoforms of platelet-derived growth factor (PDGF) were produced and purified in milligram quantities by expression of PDGF A-and B-chains in yeast cells. Structural analysis of the purified short and long variants of PDGF-AA (PDGF-AAS and PDGF-AAL) and PDGF-BB showed that they had been properly processed and assembled into dimers. PDGF-AAs and PDGF-AAL were found to bind only to the PDGF A-type receptor on human fibroblasts, with affinities of 0.1 and 0.2 uM, respectively. PDGF-BB bound to cells with A-and B-type receptors and to cells with B-type receptor only with affinities of 0.6 nM in both cases. Each fibroblast appeared to express about 4-5 times more B-type receptors than A-type receptors. The maximal mitogenic response to PDGF-BB of human fibroblasts was almost 2-fold higher than that induced by either of the two PDGF-AA forms. The three isoforms of PDGF also stimulated growth in soft agar of human fibroblasts with PDGF-BB inducing a higher maximal response.

AB - Three recombinant homodimeric isoforms of platelet-derived growth factor (PDGF) were produced and purified in milligram quantities by expression of PDGF A-and B-chains in yeast cells. Structural analysis of the purified short and long variants of PDGF-AA (PDGF-AAS and PDGF-AAL) and PDGF-BB showed that they had been properly processed and assembled into dimers. PDGF-AAs and PDGF-AAL were found to bind only to the PDGF A-type receptor on human fibroblasts, with affinities of 0.1 and 0.2 uM, respectively. PDGF-BB bound to cells with A-and B-type receptors and to cells with B-type receptor only with affinities of 0.6 nM in both cases. Each fibroblast appeared to express about 4-5 times more B-type receptors than A-type receptors. The maximal mitogenic response to PDGF-BB of human fibroblasts was almost 2-fold higher than that induced by either of the two PDGF-AA forms. The three isoforms of PDGF also stimulated growth in soft agar of human fibroblasts with PDGF-BB inducing a higher maximal response.

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JO - Growth Factors

JF - Growth Factors

SN - 0897-7194

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Expression of three recombinant homodimeric isoforms of PDGF in saccharomyces cerevisiae: Evidence for difference in receptor binding and functional activities

Abstract

Keywords

ASJC Scopus subject areas

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