G protein–dependent basal and evoked endothelial cell vWF secretion

Autor: Alexandra V. Andreeva, Xiaoping Du, David J. Visintine, Jaehyung Cho, Richard D. Minshall, Farnaz R. Bakhshi, Luiza Rusu, Osamu Kusano-Arai, Kyungho Kim, Nobuchika Suzuki, Sandra L. Haberichter, Aleksandra Stojanovic-Terpo, Takao Hamakubo, Guoquan Liu, Olga Chernaya, Hiroko Iwanari, Stephen M. Vogel, Tohru Kozasa
Rok vydání: 2014
Předmět:
congenital
hereditary
and neonatal diseases and abnormalities
G protein
Immunology
GTP-Binding Protein alpha Subunits
G12-G13
Biochemistry
Umbilical vein
Mice
Basal (phylogenetics)
Platelet Adhesiveness
Thrombin
Von Willebrand factor
Vascular Biology
hemic and lymphatic diseases
von Willebrand Factor
Human Umbilical Vein Endothelial Cells
medicine
Animals
Humans
Secretion
RNA
Small Interfering
Mice
Knockout
Hemostasis
biology
Antibodies
Monoclonal
Endothelial Cells
Thrombosis
Cell Biology
Hematology
Molecular biology
Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
Endothelial stem cell
Gene Expression Regulation
cardiovascular system
biology.protein
GTP-Binding Protein alpha Subunits
Gq-G11
rhoA GTP-Binding Protein
Protein Binding
Signal Transduction
circulatory and respiratory physiology
medicine.drug
Zdroj: Blood. 123:442-450
ISSN: 1528-0020
0006-4971
DOI: 10.1182/blood-2013-03-489351
Popis: von Willebrand factor (vWF) secretion by endothelial cells (ECs) is essential for hemostasis and thrombosis; however, the molecular mechanisms are poorly understood. Interestingly, we observed increased bleeding in EC-Gα13(-/-);Gα12(-/-) mice that could be normalized by infusion of human vWF. Blood from Gα12(-/-) mice exhibited significantly reduced vWF levels but normal vWF multimers and impaired laser-induced thrombus formation, indicating that Gα12 plays a prominent role in EC vWF secretion required for hemostasis and thrombosis. In isolated buffer-perfused mouse lungs, basal vWF levels were significantly reduced in Gα12(-/-), whereas thrombin-induced vWF secretion was defective in both EC-Gαq(-/-);Gα11(-/-) and Gα12(-/-) mice. Using siRNA in cultured human umbilical vein ECs and human pulmonary artery ECs, depletion of Gα12 and soluble N-ethylmaleimide-sensitive-fusion factor attachment protein α (α-SNAP), but not Gα13, inhibited both basal and thrombin-induced vWF secretion, whereas overexpression of activated Gα12 promoted vWF secretion. In Gαq, p115 RhoGEF, and RhoA-depleted human umbilical vein ECs, thrombin-induced vWF secretion was reduced by 40%, whereas basal secretion was unchanged. Finally, in vitro binding assays revealed that Gα12 N-terminal residues 10-15 mediated the binding of Gα12 to α-SNAP, and an engineered α-SNAP binding-domain minigene peptide blocked basal and evoked vWF secretion. Discovery of obligatory and complementary roles of Gα12 and Gαq/11 in basal vs evoked EC vWF secretion may provide promising new therapeutic strategies for treatment of thrombotic disease.
Databáze: OpenAIRE
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