Academic Journal
The bioactivity of human bone morphogenetic protein-15 is sensitive to C-terminal modification: characterization of the purified untagged processed mature region
| العنوان: | The bioactivity of human bone morphogenetic protein-15 is sensitive to C-terminal modification: characterization of the purified untagged processed mature region |
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| المؤلفون: | Pulkki, Minna M., Myllymaa, Samu, Pasternack, Arja, Lun, Stanley, Ludlow, Helen, Al-Qahtani, Ahmed, Korchynskyi, Olexandr, Groome, Nigel, Juengel, Jennifer L., Kalkkinen, Nisse, Laitinen, Mika, Ritvos, Olli, Mottershead, David G. |
| المساهمون: | Department of Bacteriology and Immunology Helsinki, Haartman Institute Helsinki, Faculty of Medecine Helsinki, University of Helsinki-University of Helsinki-Faculty of Medecine Helsinki, University of Helsinki-University of Helsinki, Oxford Brookes University, Molecular Cell Biology Department, Leiden University Medical Center (LUMC), University of Helsinki, Biotechvisions Ltd., University of Adelaide |
| المصدر: | ISSN: 0303-7207. |
| بيانات النشر: | HAL CCSD Elsevier |
| سنة النشر: | 2010 |
| المجموعة: | Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) |
| مصطلحات موضوعية: | BMP15, bone morphogenetic protein, granulosa cell, human |
| الوصف: | International audience ; Oocyte-derived bone morphogenetic protein-15 (BMP15) is critical for the regulation of mammalian fertility. Previously we have found that a C-terminal His-tag destroys the bioactivity of growth differentiation-9 (GDF9, a homolog of BMP15). In this study we found that recombinant human BMP15 is produced by HEK-293T cells in an active form, but the bioactivity is lost by C-terminal modification, specifically, fusion to a Flag tag. After purification the mature BMP15wt is active in transcriptional reporter assays specific for Smad1/5/8 in human granulosa-luteal (hGL) and COV434 granulosa tumor cells, whereas BMP15 with a carboxy-terminal Flag tag remains inactive. Using these same cell models we found that treatment with purified mature BMP15wt causes a rapid phosphorylation of Smad1. The purified BMP15wt is a potent stimulator of rat granulosa cell DNA synthesis, which could be antagonized by the BMPRII ectodomain-Fc fusion molecule, whereas the BMP15C-Flag was completely inactive. Further, the BMP15wt form is a potent stimulator of inhibin B production in hGL cells. We found that the purified BMP15wt consists of P16 and -17, both of which are post-translationally modified forms. This is the first characterization of a purified untagged human BMP15 mature region, which is stable and highly bioactive in human and rodent granulosa cells and as such is of importance for studies on human fertility. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| Relation: | hal-00654966; https://hal.archives-ouvertes.fr/hal-00654966; https://hal.archives-ouvertes.fr/hal-00654966/document; https://hal.archives-ouvertes.fr/hal-00654966/file/PEER_stage2_10.1016%252Fj.mce.2010.10.002.pdf |
| DOI: | 10.1016/j.mce.2010.10.002 |
| الاتاحة: | https://hal.archives-ouvertes.fr/hal-00654966 https://hal.archives-ouvertes.fr/hal-00654966/document https://hal.archives-ouvertes.fr/hal-00654966/file/PEER_stage2_10.1016%252Fj.mce.2010.10.002.pdf https://doi.org/10.1016/j.mce.2010.10.002 |
| Rights: | info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: | edsbas.E1C26F8 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1016/j.mce.2010.10.002 |
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