High-yield purification of platelet-derived endothelial cell growth factor: structural characterization and establishment of a specific antiserum

Miyazono, Kohei; Heldin, Carl Henrik

doi:10.1021/bi00430a042

Cited by 50 publications

(12 citation statements)

References 22 publications

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“…In general they derive from both recruited inflammatory cells as well as parenchymal. cells and include platelet-derived endothelial cell growth factor (40)(41)(42), and macrophage-derived transforming growth factor alpha (TGFa) (43,44). The relative importance of these two angiogenesis factors to the new vessel growth observed after acute lung injury remains uncertain.…”

Section: Discussionmentioning

confidence: 99%

Identification and partial characterization of angiogenesis bioactivity in the lower respiratory tract after acute lung injury.

et al. 1991

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Survival after acute lung injury (ALI) depends on prompt alveolar repair, a process frequently subverted by the development of granulation tissue within the alveolar airspace. Immunohistochemical examination of the intraalveolar granulation tissue confirmed that capillaries as well as myofibroblasts were the principal cellular constituents. We therefore hypothesized that angiogenesis factors would be present on the air-lung interface after ALI. To evaluate this hypothesis, bronchoalveolar lavage fluid from patients with ALI (a = 25) and patient controls (a = 8) was examined for angiogenesis bioactivity by its ability of induce endothelial cell migration. While lavage fluid from controls had no bioactivity, lavage fluid from 72% of patients with ALI promoted endothelial cell migration. Heparin affinity, ion exchange, and gel filtration chromatography resolved the bioactivity into at least two moieties. One appeared identical to the well characterized endothelial cell growth factor, basic fibroblast growth factor. The other was a 150-kD non-heparin binding protein that mediated endothelial cell migration and attachment in vitro, and the growth of new vessels in vivo. These data are consistent with the hypothesis that the growth of capillaries into the alveolar airspace results from angiogenesis factors present on the alveolar surface of the lung after ALI. (J. Clin. Invest. 1991. 88:1386-1395

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Section: Discussionmentioning

confidence: 99%

Identification and partial characterization of angiogenesis bioactivity in the lower respiratory tract after acute lung injury.

et al. 1991

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“…Platelet-derived endothelial cell growth factor (PD-ECGF) was originally isolated from platelets through its unique mitogenic activity on endothelial cells (Miyazono et al, 1987;1989). It has since been shown to be homologous to human thymidine phosphorylase and a product of the same gene (Furukawa et al, 1992).…”

mentioning

confidence: 99%

Increased sensitivity to the prodrug 5'-deoxy-5-fluorouridine and modulation of 5-fluoro-2'-deoxyuridine sensitivity in MCF-7 cells transfected with thymidine phosphorylase

Patterson

Zhang²,

Moghaddam³

et al. 1995

Br J Cancer

116

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Summary Platelet-derived endothelial cell growth factor (PD-ECGF) is identical to human thymidine phosphorylase (dThdPase). The human MCF-7 breast cancer cell line was transfected with the dThdPase cDNA and expressed a 45 kDa protein that was detected with anti-dThdPase antibody. Cell lysates possessed elevated dThdPase activity and cells had up to 165-fold increased sensitivity to the prodrug 5'-deoxy-5-fluorouridine (5'-DFUR) in vitro. Sensitivity to 5-fluorouracil (5-FU) and 5-fluoro-2'-deoxyuridine (5-FUdR) was unchanged. Recombinant dThdPase was shown to catalyse directly the phosphorolytic cleavage of 5'-DFUR to 5-FU. Exogenous thymidine (dThd) reversed the toxicity of 5-FUdR on the parental line (1 JAM dThd increased the IC50 value 1000-fold), but the dThd rescue was substantially modulated in the dThdPaseexpressing clone 4 (1 gM dThd raised the IC50 value 3-fold). We observed a substantial 'bystander' killing effect when small proportions of dThdPase-expressing cells were mixed with parental MCF-7 cells. dThdPase activity was on average 27-fold higher in breast tumours than in normal breast. The levels in wild-type MCF-7 are similar to the low end of the tumour expression. Thus, in some tumours resistance to 5'-DFUR therapy could be due to low dThdPase activity, and transfection to raise the dThdPase. levels within the broad tumour range or above it should markedly enhance sensitivity to the prodrug. These results confirm that dThdPase is a major pathway in the metabolic activation of 5'-DFUR, and the bystander effect suggests that this may be a suitable enzyme for gene therapy-directed enzyme/prodrug activation therapy.

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“…It is a 45-kDa protein which, in contrast to angiogenic factors of the fibroblast growth factor (FGF) family (4), does not stimulate the growth of fibroblasts. PD-ECGF has been purified from human platelet lysate and seems to be the only endothelial cell mitogen in platelets (5). cDNA for PD-ECGF was recently cloned (3); the predicted amino acid sequence has no striking homology with that of previously known proteins.…”

mentioning

confidence: 99%

Production of platelet-derived endothelial cell growth factor by normal and transformed human cells in culture.

Usuki

Heldin

Miyazono

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

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Platelet-derived endothelial cell growth factor (PD-ECGF) is a 45-kDa endothelial cell mitogen which has angiogenic properties in vivo. We report here that human foreskin fibroblasts, a human squamous cell carcinoma cell line, and 2 out of the 3 human thyroid carcinoma cell lines investigated produce PD-ECGF, whereas 21 other cell lines examined do not. The positive cell lines contained a 1.8-kilobase PD-ECGF mRNA, and a 45-kDa protein could be demonstrated in lysates of the cell lines by immunoblotting and immunoprecipitation using a specific antiserum against PD-

show abstract

High-yield purification of platelet-derived endothelial cell growth factor: structural characterization and establishment of a specific antiserum

Cited by 50 publications

References 22 publications

Identification and partial characterization of angiogenesis bioactivity in the lower respiratory tract after acute lung injury.

Identification and partial characterization of angiogenesis bioactivity in the lower respiratory tract after acute lung injury.

Increased sensitivity to the prodrug 5'-deoxy-5-fluorouridine and modulation of 5-fluoro-2'-deoxyuridine sensitivity in MCF-7 cells transfected with thymidine phosphorylase

Production of platelet-derived endothelial cell growth factor by normal and transformed human cells in culture.

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