cGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interaction
Abstract Background The nitric oxide-soluble guanylate cyclase-cyclic guanosine monophosphate (NO-sGC-cGMP) signaling pathway, plays a critical role in the pathogenesis of pulmonary arterial hypertension (PAH); however, its exact molecular mechanism remains undefined. Methods Biotin-cGMP pull-down a...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-10-01
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| Series: | Respiratory Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12931-018-0903-z |
| _version_ | 1819265171876478976 |
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| author | Lihui Zou Junhua Zhang Jingli Han Wenqing Li Fei Su Xiaomao Xu Zhenguo Zhai Fei Xiao |
| author_facet | Lihui Zou Junhua Zhang Jingli Han Wenqing Li Fei Su Xiaomao Xu Zhenguo Zhai Fei Xiao |
| author_sort | Lihui Zou |
| collection | DOAJ |
| description | Abstract Background The nitric oxide-soluble guanylate cyclase-cyclic guanosine monophosphate (NO-sGC-cGMP) signaling pathway, plays a critical role in the pathogenesis of pulmonary arterial hypertension (PAH); however, its exact molecular mechanism remains undefined. Methods Biotin-cGMP pull-down assay was performed to search for proteins regulated by cGMP. The interaction between cGMP and tropomyosin was analyzed with antibody dependent pull-down in vivo. Tropomyosin fragments were constructed to explore the tropomyosin-cGMP binding sites. The expression level and subcellular localization of tropomyosin were detected with Real-time PCR, Western blot and immunofluorescence assay after the 8-Br-cGMP treatment. Finally, isothermal titration calorimetry (ITC) was utilized to detect the binding affinity of actin-tropomyosin-myosin in the existence of cGMP-tropomyosin interaction. Results cGMP interacted with tropomyosin. Isoform 4 of TPM1 gene was identified as the only isoform expressed in the human pulmonary artery smooth muscle cells (HPASMCs). The region of 68-208aa of tropomyosin was necessary for the interaction between tropomyosin and cGMP. The expression level and subcellular localization of tropomyosin showed no change after the stimulation of NO-sGC-cGMP pathway. However, cGMP-tropomyosin interaction decreased the affinity of tropomyosin to actin. Conclusions We elucidate the downstream signal pathway of NO-sGC-cGMP. This work will contribute to the detection of innovative targeted agents and provide novel insights into the development of new therapies for PAH. |
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| format | Article |
| id | doaj.art-5c1027d88ddb461189607cf646e414d0 |
| institution | Directory Open Access Journal |
| issn | 1465-993X |
| language | English |
| last_indexed | 2024-12-23T20:41:09Z |
| publishDate | 2018-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Respiratory Research |
| spelling | doaj.art-5c1027d88ddb461189607cf646e414d02022-12-21T17:31:55ZengBMCRespiratory Research1465-993X2018-10-011911910.1186/s12931-018-0903-zcGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interactionLihui Zou0Junhua Zhang1Jingli Han2Wenqing Li3Fei Su4Xiaomao Xu5Zhenguo Zhai6Fei Xiao7The MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of GerontologyThe MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of GerontologyThe MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of GerontologyThe MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of GerontologyDepartment of Pathology, Beijing Hospital, National Center of GerontologyDepartment of Respiratory and Critical Care Medicine, Beijing Hospital, National Center of GerontologyDepartment of Respiratory and Critical Care Medicine, China-Japan Friendship HospitalThe MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of GerontologyAbstract Background The nitric oxide-soluble guanylate cyclase-cyclic guanosine monophosphate (NO-sGC-cGMP) signaling pathway, plays a critical role in the pathogenesis of pulmonary arterial hypertension (PAH); however, its exact molecular mechanism remains undefined. Methods Biotin-cGMP pull-down assay was performed to search for proteins regulated by cGMP. The interaction between cGMP and tropomyosin was analyzed with antibody dependent pull-down in vivo. Tropomyosin fragments were constructed to explore the tropomyosin-cGMP binding sites. The expression level and subcellular localization of tropomyosin were detected with Real-time PCR, Western blot and immunofluorescence assay after the 8-Br-cGMP treatment. Finally, isothermal titration calorimetry (ITC) was utilized to detect the binding affinity of actin-tropomyosin-myosin in the existence of cGMP-tropomyosin interaction. Results cGMP interacted with tropomyosin. Isoform 4 of TPM1 gene was identified as the only isoform expressed in the human pulmonary artery smooth muscle cells (HPASMCs). The region of 68-208aa of tropomyosin was necessary for the interaction between tropomyosin and cGMP. The expression level and subcellular localization of tropomyosin showed no change after the stimulation of NO-sGC-cGMP pathway. However, cGMP-tropomyosin interaction decreased the affinity of tropomyosin to actin. Conclusions We elucidate the downstream signal pathway of NO-sGC-cGMP. This work will contribute to the detection of innovative targeted agents and provide novel insights into the development of new therapies for PAH.http://link.springer.com/article/10.1186/s12931-018-0903-zCyclic guanosine monophosphateTropomyosinActinMyosinInteraction |
| spellingShingle | Lihui Zou Junhua Zhang Jingli Han Wenqing Li Fei Su Xiaomao Xu Zhenguo Zhai Fei Xiao cGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interaction Respiratory Research Cyclic guanosine monophosphate Tropomyosin Actin Myosin Interaction |
| title | cGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interaction |
| title_full | cGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interaction |
| title_fullStr | cGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interaction |
| title_full_unstemmed | cGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interaction |
| title_short | cGMP interacts with tropomyosin and downregulates actin-tropomyosin-myosin complex interaction |
| title_sort | cgmp interacts with tropomyosin and downregulates actin tropomyosin myosin complex interaction |
| topic | Cyclic guanosine monophosphate Tropomyosin Actin Myosin Interaction |
| url | http://link.springer.com/article/10.1186/s12931-018-0903-z |
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