Proteome Science | |
Proteomic analysis of secretagogue-stimulated neutrophils implicates a role for actin and actin-interacting proteins in Rac2-mediated granule exocytosis | |
Paige Lacy2  Danny V Chao2  John D Kim2  Troy Mitchell1  Andrea N Lo2  Gary Eitzen1  | |
[1] Department of Cell Biology, University of Alberta, Edmonton, AB, Canada;Pulmonary Research Group, Department of Medicine, University of Alberta, Edmonton, AB, Canada | |
关键词: coronin; actin; GTPase; Rac2; exocytosis; degranulation; neutrophil; | |
Others : 820092 DOI : 10.1186/1477-5956-9-70 |
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received in 2011-07-24, accepted in 2011-11-14, 发布年份 2011 | |
【 摘 要 】
Background
Neutrophils are abundant leukocytes that play a primary role in defence against pathogens. Neutrophils enter sites of infection where they eliminate pathogens via phagocytosis and the release of antimicrobial mediators via degranulation. Rho GTPases, particularly Rac2, play a key role in neutrophil degranulation. The purpose of this study was to identify Rac2-dependent changes in protein abundance in stimulated neutrophils.
Methods
We performed a proteomic analysis on secretagogue-stimulated bone marrow neutrophils that were isolated from wild-type and Rac2-/- mice. Protein abundance was analyzed by 2-dimensional SDS-PAGE of fluorescently labelled samples which allowed the detection ~3500 proteins.
Results
We identified 22 proteins that showed significant changes in abundance after secretagogue-stimulation of wild-type neutrophils, which did not occur in neutrophils isolated from Rac2-/- mice. As expected, the abundance of several granule proteins was reduced in wild-type cells; this did not occur in Rac2-/- neutrophils which confirms the requirement for Rac2 in degranulation. We also found changes in abundance of many actin remodelling proteins including coronin-1A, β-actin and the F-actin capping protein, (CapZ-β). Coronin-1A showed elevated levels of several isoforms after stimulation of neutrophils from wild-type, but not from Rac2-/- mice. These isoforms were immunoreactive with anti-phospho-threonine antibodies, suggesting that neutrophil stimulation triggers a Rac2-dependent kinase cascade that results in the phosphorylation of coronin-1A.
Conclusion
The control of Rac2-mediated degranulation in neutrophils likely functions through actin remodelling via activation of several actin-binding proteins. We found coronin-1A to be a novel downstream effector protein of this pathway that is threonine phosphorylated in response to secretagogue stimulation.
【 授权许可】
2011 Eitzen et al; licensee BioMed Central Ltd.
【 预 览 】
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