Analysis of Small Latent Transforming Growth Factor-β Complex Formation and Dissociation by Surface Plasmon Resonance

Bailly, Sabine; Brand, Céline; Chambaz, E.M.; Feige, Jean‐Jacques

doi:10.1074/jbc.272.26.16329

Cited by 34 publications

(24 citation statements)

References 25 publications

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“…The tripeptidic motif RFK, present within the type I repeats of TSP1ang, was previously reported to have the capacity to activate latent TGFb in long-term cellular assays (Murphy-Ullrich and Poczatek, 2000). Whether this results from a direct or an indirect effect is still an open question, since delicate molecular techniques such as surface plasmon resonance using purified molecules did not allow to visualize a direct interaction between TSPs and latent TFb (Bailly et al, 1997). Recent studies have suggested that an indirect mechanism could be responsible for this activation (Grainger and Frow, 2000;Harpel et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

Expression of the thrombospondin 1 fragment 167–569 in C6 glioma cells stimulates tumorigenicity despite reduced neovascularization

et al. 2004

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Gliomas are among the most malignant and most highly vascularized human tumors. We studied the therapeutic action of an angiostatic fragment of human thrombospondin 1 (named TSP1ang) on experimental glioma tumor growth. TSP1ang (enclosing amino acids 167-569) comprised the procollagen-homology domain and the three type I repeats of the original molecule. C6 glioma cells that stably express TSP1ang were generated, and their rate of in vitro growth did not appear to differ from that of C6 cells transfected with an empty plasmid. TSP1ang-expressing C6 cells were then injected either subcutaneously or intracerebrally into nude mice. The resulting tumors appeared to be less vascularized, but unexpectedly started to grow earlier and had a much more invasive phenotype than tumors derived from control C6 cells. They were also much more aggressive, since the mice bearing intracerebral TSP1ang-expressing tumor cells died before day 19 post-implantation, whereas all mice bearing control C6 tumors were alive at this time point. These results indicate that careful attention should be paid at designing smaller fragments from the multimodular angiostatic molecule TSP1 since, as observed in this study, it may unmask protumorigenic properties that counteract its antiangiogenic activity.

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Section: Discussionmentioning

confidence: 99%

Expression of the thrombospondin 1 fragment 167–569 in C6 glioma cells stimulates tumorigenicity despite reduced neovascularization

et al. 2004

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“…We ultimately chose to evaluate rLAP as a possible inhibitor because this molecule is as TGF-␤1 specific as anti-TGF-␤1 and binds to TGF-␤1 with at least as high an affinity as anti-TGF-␤1 (24). This substance could conceivably block TGF-␤1 suppressor activity by either of two mechanisms: 1) by combining with newly released active TGF-␤1 and converting it back into a latent form; and 2) by competitive inhibition of latent TGF-␤1 (LAP) for potential binding sites at which conversion of latent TGF-␤1 to an active form normally occurs.…”

Section: Cd25mentioning

confidence: 99%

TGF-β1 Plays an Important Role in the Mechanism of CD4+CD25+ Regulatory T Cell Activity in Both Humans and Mice

Nakamura

Kitani

Fuss

et al. 2004

The Journal of Immunology

578

463

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In previous studies, we have shown that murine CD4+CD25+ regulatory T cells produce high levels of TGF-β1 in a cell surface and/or secreted form, and blockade of such TGF-β1 by anti-TGF-β curtails the ability of these cells to suppress CD25− T cell proliferation and B cell Ig production in in vitro suppressor assays. In further support for the role of TGF-β1 in suppression by CD4+CD25+ T cells, we show in this study that another TGF-β1-blocking molecule, recombinant latency-associated peptide of TGF-β1 (rLAP), also reverses suppression by mouse CD4+CD25+ T cells as well as their human counterparts, CD4+CD25high T cells. In addition, we show that CD25− T cells exposed to CD4+CD25+ T cells in vitro manifest activation of Smad-2 and induction of CD103, the latter a TGF-β-inducible surface integrin. In further studies, we show that while CD4+CD25+ T cells from TGF-β1-deficient mice can suppress CD25− T cell proliferation in vitro, these cells do not protect recipient mice from colitis in the SCID transfer model in vivo, and, in addition, CD4+LAP+, but not CD4+LAP− T cells from normal mice protect recipient mice from colitis in this model. Together, these studies demonstrate that TGF-β1 produced by CD4+CD25+ T cells is involved in the suppressor activity of these cells, particularly in their ability to regulate intestinal inflammation.

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“…The isoforms have similar biological activities but exhibit differences in potency depending on the target cell examined (7,8). TGF-␤ is produced by virtually all cell types as inactive precursors (1,9) and receptors for TGF-␤ are universally distributed throughout the body. The inactive precursor is first cleaved into a latent complex, which upon activation by acidification represents the regulation step of the signaling process of TGF-␤ (7, 10).…”

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confidence: 99%

Conformation and Self-association of Human Recombinant Transforming Growth Factor-β3 in Aqueous Solutions

Pellaud

Schote

Arvinte

et al. 1999

Journal of Biological Chemistry

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The transforming growth factors-␤ (TGF-␤) are important regulatory peptides for cell growth and differentiation with therapeutic potential for wound healing. Among the several TGF-␤ isoforms TGF-␤3 has a particularly low solubility at physiological pH and easily forms aggregates. A spectroscopic structural analysis of TGF-␤3 in solution has thus been difficult. In this study, circular dichroism spectroscopy was used to determine the secondary structural elements of TGF-␤3. In addition, the aggregation of TGF-␤3 was investigated systematically as a function of pH and salt concentration using a rapid screening method. Sedimentation equilibrium and sedimentation velocity analysis revealed that TGF-␤3 exists predominantly in two major forms: (i) monomers in solution at low pH and (ii) large precipitating aggregates at physiological pH. Under acidic conditions (pH < 3.8) the protein was not aggregated. At pH ϳ3.9, a monomer i dimer equilibrium could be detected that transformed into larger aggregates at pH > 4.1. Aggregation was pronounced in the pH range of 4.3 < pH < 9.8 with the aggregation maximum between pH 6.5 and 8.5. The aggregation process was accompanied by a structural change of the protein. The CD spectra were characterized by an isodichroic point at 209.5 nm indicating a two-state equilibrium between TGF-␤3 dissolved in solution and aggregated TGF-␤3. Aggregated TGF-␤3 showed a higher ␤-sheet content and lower ␤-turn and random coil contributions compared with monomeric TGF-␤3. Both the solution structure and the aggregate structure of TGF-␤3 were different from the crystal structure. This was in contrast to TGF-␤2, which showed very similar crystal and solution structures. Under alkaline conditions (pH > 9.8) the turbidity disappeared and a further conformational change was induced. The pH dependence of the TGF-␤3 conformation in solution in the range of 2.3 < pH < 11.0 was reversible. Aggregation of TGF-␤3 was, furthermore, influenced by the presence of salt. For pH > 3.8 the addition of salt greatly enhanced the tendency to aggregate, even in the very basic domain. Under physiological conditions (pH 7.4, c NaCl ‫؍‬ 164 mM) TGF-␤3 has almost the highest tendency to aggregate and will remain in solution only at nanomolar concentrations.

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Analysis of Small Latent Transforming Growth Factor-β Complex Formation and Dissociation by Surface Plasmon Resonance

Cited by 34 publications

References 25 publications

Expression of the thrombospondin 1 fragment 167–569 in C6 glioma cells stimulates tumorigenicity despite reduced neovascularization

Expression of the thrombospondin 1 fragment 167–569 in C6 glioma cells stimulates tumorigenicity despite reduced neovascularization

TGF-β1 Plays an Important Role in the Mechanism of CD4+CD25+ Regulatory T Cell Activity in Both Humans and Mice

Conformation and Self-association of Human Recombinant Transforming Growth Factor-β3 in Aqueous Solutions

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