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TGF-Βsuperfamily R&D Systems Tools for Cell Biology Research™ Nodal BMP-8 TGF-b Superfamily BMP-7 BMP-5 GDF-10 GDF-1 BMP-11 BMP-3 BMP-4 BMP-6 GDF-3 GDF-8 BMP-2 BMP-9 Activin/Inhibin βC GDF-7 Activin/Inhibin βE BMP-10 Activin/Inhibin βB GDF-5 GDF-6 Activin/Inhibin βA BMP-15 GDF-9 MIS TGF-β1 Inhibin α TGF-β2 TGF-β3 Neurturin GDNF Lefty-B Lefty-A Persephin Artemin The TGF-b Superfamily The transforming growth factor beta (TGF-b) superfamily contains highly pleiotropic molecules that encompass diverse functions during embryo- genesis and adult tissue homeostasis. Mammalian members of the TGF-b superfamily include TGF-b1,-b2,-b3, Activins, Inhibins, BMPs (bone mor- phogenetic proteins), GDFs (growth differentiation factors), GDNFs (glial derived neurotrophic factors), Nodal, Lefty, and MIS (Müllerian inhibiting substance). Sequence profile analysis of the TGF-b superfamily members indicate that they have a close structural similarity, sharing approximately 25-40% amino acid sequence identity. TGF-b ligands are initially synthesized as precursor proteins that undergo proteolytic cleavage. The mature segments form active ligand dimers via a disulfide-rich core consisting of the characteristic “cysteine knot”. Although homodimers are considered the standard form, there are natural heterodimers with biological activity as well. Chordin LAP Noggin TGF-β dimers Small latent TGF-β BMP dimers Large latent TGF-β Growth BMPR-II Factor R TGF-β RIII RGM Receptor LTBP TGF-β RII ECM BMPR-I TGF-β RI P P SARA XIAP RAS Smad6/7 Smad7 TAB1 ERK P P Smad2/3 Smad1/5/8 Smad4 TAK1 Smurf Smad4 Smad1/5/8 P P Smad4 P P Smurf Smad2/3 Smad4 MAPK Signaling MAPK/ERK (p38, JNK) JUN FOS Smad4 CoR Smad P Proliferation, Apoptosis, Morphogenesis, CoA Differentiation, ECM Remodeling, Migration Transcription factors CoR = Co-Repressors CoA = Co-Activators TGF-b ligands bind to heteromeric receptor complexes with serine/threonine kinase domains. Upon ligand binding, the type II receptor activates the type I receptor by phosphorylation of the GS region. The type I receptor phosphorylates a receptor-activated Smad (R-Smad) protein. Phos- phorylated R-Smads form a trimeric complex with the common Smad4, and this complex moves into the nucleus to interact with transcriptional co-activators or co-repressors. This signaling cascade that ultimately alters gene transcription and cellular responses is regulated at several levels. Co-receptors may associate with ligands and act as obligate cofactors (such as TGF-b RIII for TGF-b2) enabling signal transduction, or act to enhance signaling by increasing sensitivity to low ligand concentrations (such as RGM for BMP signaling). The particular type I-type II receptor combination, the activated Smad pathway, and the cell-specific transcription factors also offer diversity in responses. Contributions of accessory proteins such as soluble or membrane-bound regulators, and the use of Smad-independent signaling pathways further fine-tune TGF-b superfamily signaling and allow for disparate activities observed in response to TGF-b ligands in different contexts. www.RnDSystems.com TGF-b-Related Products ANTIBODIES ANTIBODIES ELISAs/ ELISAs/ MOLECULE PROTEINS ASSAYS MOLECULE PROTEINS ASSAYS POLYCLONAL MONOCLONAL LABELED POLYCLONAL MONOCLONAL LABELED Activin A H M R H M R H M R H M R H M R CRIM1 H M H H H Activin AB, AC H Cripto H M H M H M H M H Cryptic H H M H M H M Activin B H H CV-2 H M H M H M H M Activin C H M H M DAN H M H M H M H M H M Activin RIA/ALK-2 H H H H Decapentaplegic D D D Activin RIB/ALK-4 H M H M H M H Decorin H M H M H M H M H M Activin RIIA H H H H Dermatopontin H Activin RIIB H M H H H Endoglin/CD105 H M P H M H M H M H M ALK-1 H M H M H M H M M FLRG H M H M H M H M H ALK-7 R R R R Follistatin H M H H H H Artemin H M H M H M H M Follistatin-like 1 H M H M R H M BAMBI/NMA H M H M Follistatin-like 4 H H H BMP-1/PCP H H H H GASP-1/ BMP-2 H Z H H Z H H M R WFIKKNRP H H H H H BMP-2/7 GASP-2/WFIKKN H H H H H Heterodimer H H GDF-1 M M BMP-3 H H H H GDF-3 H M M M M BMP-3b/GDF-10 H H GDF-5 M M M M BMP-4 H M Z H H Z H H GDF-6 M BMP-4/7 Heterodimer H GDF-7/BMP-12 M M M BMP-5 H M H H H H GDF-8 Propeptide M M H M R M BMP-6 H M H H H H GDF-8/Myostatin H M R H M R H M R H M R BMP-7 H M H H H H GDF-9 M M M BMP-8 H H H GDF-11/BMP-11 H BMP-9 H M H H H H GDF-15 H H H H H BMP-10 H M H H H GDNF H R H R H H R H BMP-15/GDF-9B H H H M H GFRa-1 H R H R H R H R BMPR-IA/ALK-3 H M H H H GFRa-2 H M H M H H M BMPR-IB/ALK-6 H M H H M H GFRa-3 H M H M H H M BMPR-II H H H H GFRa-4 H M H M M Caronte Ch Ch Ch Ch Gremlin H M M M CD109 H H H H Lefty H M H M H M H M Cerberus 1 M H M H M H M MIS/AMH H H R H M R Chordin M M M M M MIS RII H R H R H Chordin-Like 1 H H H H Neurturin H M H M H H M Chordin-Like 2 M H M H M H M Nodal H M M M M COCO H M H M H M H M H Noggin H M M M M For a complete & up-to-date listing, visit www.RnDSystems.com/go/TGFbeta For research use only. Not for use in diagnostic procedures. ANTIBODIES ELISAs/ MOLECULE PROTEINS ASSAYS POLYCLONAL MONOCLONAL LABELED NOMO H Persephin H M H M H M H PP2Ca/PPM1A H H M R PRDC M M M M M Ret H M H M H M H M H BMP-15/GDF-9b in Mouse Ovary. Bone Morphoge- Twisted Gastrulation (TSG) Expression in netic Protein-15 (BMP-15) was detected in a frozen section Embryonic Mouse Ribs. Detection of TSG in a Phospho-Ret H (Y905) of adult mouse ovary using Human BMP-15 Monoclonal cryostat tissue section of mouse embryonic (15 d.p.c.) rib Antibody (Catalog # MAB2925). Cells were stained using cartilage primordium using Mouse Twisted Gastrulation Phospho-Ret (Y1062) H NorthernLights-557-conjugated Anti-Rat IgG Secondary Antigen Affinity-purified Polyclonal Antibody (Catalog # Antibody (red; Catalog # NL013). Nuclei were counter- AF756). Tissues were stained using Rhodamine Red™ X- SKI H stained with DAPI (blue). conjugated anti-goat secondary antibody (red) and coun- terstained with Fluoro Nissl Green (green). Smad1, 5, 8 H Smad2 H M H H Smad3 H M H H H GDNF Antibody (µg/mL) .01 0.1 1 Phospho-Smad3 0.32 (S423/S425) H Smad4 H H H H 0.28 Smad7 H M R SOST/Sclerostin H M H M H M H M 0.24 Relative Cell Number Cell Relative TGF-b1 H P Ms Ms Ms H M R Ca P 0.20 LAP (TGF-b1) H H H H M 0 0.1 1 10 BMPR-II in Mouse Embryo. BMPR-II was detected in a Recombinant GDNF (ng/mL) frozen section of E15 mouse ribs using Human BMPR-II Latent TGF-b1 H H Antigen Affinity-purified Polyclonal Antibody (Catalog # GDNF-induced Neuronal Survival and AF811; green). TGF-b1.2 H Ms Antibody Neutralization. Chick DRG neuron sur- vival increases in response to Recombinant Human GDNF TGF-b2 H P Ms Ms Ms H (Catalog # 212-GD) in a dose-dependent manner (blue line) as measured with the MTT Cell Proliferation/Viability Assay TGF-b3 H Ms Ms Ms H 80 (Catalog # TA5355). Neuronal survival elicited by 10 ng/mL 70 TGF-b5 A Ms GDNF is neutralized (green line) by increasing concentra- tions of Human/Rat GDNF Antigen Affinity-purified Poly- 60 Latent TGF-b bp1 H H clonal Antibody (Catalog # AF-212-NA). 50 Latent TGF-b bp2 H 40 Latent TGF-b bp4 M 30 TGF-b RI/ALK-5 M H M H M H M Number Cell Relative 20 M.W. Reduced Non-Reduced TGF-b RII H M H M H M H M H (kDa) 10 TGF-b RIIb H H 0 0 1 2 3 4 29 < BMP-2/7 10 10 10 10 10 TGF-b RIII H M H M H H M H Heterodimer TGF-β-1, -β2, -β3 TMEFF1/ 21 Tomoregulin-1 H M H M 18.4 Detection of TGF-b in PC-3 cells using TSG M M M M < BMP-7 Flow Cytometry.
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