Rho/Rac/Cdc42 Activator I

Rho/Rac/Cdc42 Activator I
$0.00

Product Uses Include

  • Control for Rho, Rac and Cdc42 pathway activation

  • Study the effects of Rho family small G-protein activation on other signaling pathways

  • Study the cell-type specific cross-talk between signaling pathways for Rho, Rac and Cdc42

  • Study the effects of Rho family small G-protein activation on the re-arrangement of the actin cytoskeleton

    G-Switch-Logo-_White-BG_

    The active site of CN04 is based on the catalytic domain of the bacterial cytotoxic necrotizing factor (CNF) toxins. The catalytic domain is covalently attached to a proprietary cell penetrating moiety. CN04 directly activates Rho GTPase isoforms by deamidating glutamine-63 of Rho and glutamine-61 of Rac and Cdc42 in their respective Switch II regions (1,2). This modification converts glutamine-63 to glutamate, which blocks intrinsic and GAP-stimulated GTPase activity, resulting in constitutively active endogenous Rho, Rac and Cdc42 (3). CN04 robustly increases the level of GTP-bound RhoA, Rac1 and Cdc42 within 2-4 h after addition to the culture medium. CN04 can be used when a direct activator of Rho family proteins is required rather than a classic indirect activator (e.g., LPA, EGF, Bradykinin and Sphingosine-1-phosphate) that concomitantly activate other signaling pathways (e.g., Ras, PI3K and PLC).

     

    References

    1. Lerm M., et al. 1999. Deamidation of Cdc42 and Rac by Escherichia coli cytotoxic necrotizing factor 1: activation of c-Jun N-terminal kinase in HeLa cells. Infection and immunity. 67, 496-503.


    2. Schmidt G., et al. 1997. Gln 63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor-1. Nature. 387, 725-729.


    3. Flatau G., et al. 1997. Toxin-induced activation of the G protein p21 Rho by deamidation of glutamine. Nature. 387, 729-733.

    Above : Rac activation in Swiss 3T3 cells. F-actin is visualized with fluorescent green phalloidin staining (Cat.# PHDG1) and nuclear blue DNA staining with Dapi. Cells were activated with Cat. # CN04 (right).

    For product Datasheets and MSDSs please click on the PDF links below.   For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

    AuthorTitleJournalYearArticle Link
    Nath, Anu S. et al.Modulation of the cell membrane lipid milieu by peroxisomal β-oxidation induces Rho1 signaling to trigger inflammatory responsesCell Reports2022ISSN 2211-1247
    Jayabal, Panneerselvam et al.NELL2-cdc42 signaling regulates BAF complexes and Ewing sarcoma cell growthCell Reports2021ISSN 2211-1247
    Merlini, Mario et al.Microglial Gi-dependent dynamics regulate brain network hyperexcitabilityNature Neuroscience2021ISSN 1546-1726
    Rong, Zhouyi et al.Activation of FAK/Rac1/Cdc42-GTPase signaling ameliorates impaired microglial migration response to Aβ42 in triggering receptor expressed on myeloid cells 2 loss-of-function murine modelsFASEB Journal2020ISSN 1530-6860
    Chen, Zihang et al.Distinct roles of srGAP3-Rac1 in the initiation and maintenance phases of neuropathic pain induced by paclitaxelThe Journal of physiology2020ISSN 1469--7793
    Larribère, Lionel et al.NF1-RAC1 axis regulates migration of the melanocytic lineageTranslational Oncology2020ISSN 1936-5233
    Andrews, Madeline G. et al.Mtor signaling regulates the morphology and migration of outer radial glia in developing human cortexeLife2020ISSN 2050-084X
    Lundin, Vanessa et al.YAP Regulates Hematopoietic Stem Cell Formation in Response to the Biomechanical Forces of Blood FlowDevelopmental Cell2020ISSN 1878-1551
    Matsuda, Jun et al.ARHGEF7 (β-PIX) is required for the maintenance of podocyte architecture and glomerular functionJournal of the American Society of Nephrology2020ISSN 1533-3450
    García-Ponce, Alexander et al.Epac1 Is Crucial for Maintenance of Endothelial Barrier Function through A Mechanism Partly Independent of Rac1Cells2020ISSN 2073-4409
    Kolyvushko, Oleksandr et al.Equine alphaherpesviruses require activation of the small gtpases rac1 and cdc42 for intracellular transportMicroorganisms2020ISSN 2076-2607
    Rom, Slava et al.Author Correction: Hyperglycemia and advanced glycation end products disrupt BBB and promote occludin and claudin-5 protein secretion on extracellular microvesicles (Scientific Reports, (2020), 10, 1, (7274), 10.1038/s41598-020-64349-x)Scientific Reports2020ISSN 2045-2322
    Costa, Joana Freitas et al.The Role of Rac GTPase in Dendritic Spine Morphogenesis and MemoryFrontiers in Synaptic Neuroscience2020ISSN 1663-3563
    Li, Ling Zhu et al.Rac1 Modulates Excitatory Synaptic Transmission in Mouse Retinal Ganglion CellsNeuroscience Bulletin2019ISSN 1995-8218
    Rizzi, Caterina et al.NGF steers microglia toward a neuroprotective phenotypeGlia2018ISSN 1098-1136
    Jackson, Edwin K. et al.8-aminoguanine induces diuresis, natriuresis, and glucosuria by inhibiting purine nucleoside phosphorylase and reduces potassium excretion by inhibiting Rac1Journal of the American Heart Association2018ISSN 2047-9980
    Kimura-Yoshida, Chiharu et al.Cytoplasmic localization of GRHL3 upon epidermal differentiation triggers cell shape change for epithelial morphogenesisNature Communications2018ISSN 2041-1723
    Robins, Richard et al.Rac1 activation in podocytes induces the spectrum of nephrotic syndromeKidney International2017ISSN 1523-1755
    Jones, Eleanor L. et al.Dendritic Cell Migration and Antigen Presentation Are Coordinated by the Opposing Functions of the Tetraspanins CD82 and CD37The Journal of Immunology2016ISSN 0022--1767
    Jiang, Lizhu et al.Inhibition of Rac1 Activity in the Hippocampus Impairs the Forgetting of Contextual Fear MemoryMolecular Neurobiology2016ISSN 1559-1182
    Rom, Slava et al.PARP inhibition in leukocytes diminishes inflammation via effects on integrins/cytoskeleton and protects the blood-brain barrierJournal of Neuroinflammation2016ISSN 1742-2094
    Reinardy, Jessica L. et al.Phosphorylation of threonine 794 on Tie1 by Rac1/PAK1 reveals a novel angiogenesis regulatory pathwayPLoS ONE2015ISSN 1932-6203
    Rom, Slava et al.Poly(ADP-ribose) polymerase-1 inhibition in brain endothelium protects the blood-brain barrier under physiologic and neuroinflammatory conditionsJournal of Cerebral Blood Flow and Metabolism2015ISSN 1559-7016
    Staiculescu, Marius C. et al.Prolonged vasoconstriction of resistance arteries involves vascular smooth muscle actin polymerization leading to inward remodellingCardiovascular Research2013ISSN 0008-6363
    Rom, Slava et al.Selective activation of cannabinoid receptor 2 in leukocytes suppresses their engagement of the brain endothelium and protects the blood-brain barrierAmerican Journal of Pathology2013ISSN 0002-9440
    Rom, Slava et al.Glycogen synthase kinase 3β inhibition prevents monocyte migration across brain endothelial cells via Rac1-GTPase suppression and down-regulation of active integrin conformationAmerican Journal of Pathology2012ISSN 0002-9440
    Schmidt, Gudula et al.Gln 63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor-1Nature 1997 387:66341997ISSN 1476--4687
    Flatau, Gilles et al.Toxin-induced activation of the G protein p21 Rho by deamidation of glutamineNature1997ISSN 0028--0836

    Question 1:  Can the direct Rho/Rac/Cdc42 activator CN04 be used with cells growing in culture?

    Answer 1:  Yes, CN04 is specifically designed to be used as a Rho/Rac/Cdc42 activator with cultured cells.  The active site of CN04 is based on the catalytic domain of the bacterial cytotoxic necrotizing factor (CNF) toxins.  The catalytic domain of CN04 is covalently attached to a proprietary cell penetrating moiety.  Upon entry into the cell, CN04 directly activates Rho GTPase isoforms by deamidating glutamine-63 of Rho and glutamine-61 of Rac and Cdc42 in their respective Switch II regions. This modification converts glutamine-63 to glutamate, which blocks intrinsic and GAP-stimulated GTPase activity, resulting in constitutively active endogenous Rho, Rac and Cdc42.  CN04 robustly increases the level of GTP-bound RhoA, Rac1 and Cdc42 within 2-4 h after addition to the culture medium.

     

    Question 2:  How can I assess whether Rho, Rac and/or Cdc42 activity is changing in my cells following CN04 treatment? 

    Answer 2:  There are multiple ways to measure changes in Rho, Rac and Cdc42 activity.  To visualize a change in a cell’s cytoskeleton mediated by Rho family proteins, we recommend examining stress fiber formation and edge ruffling with fluorescently-labeled phalloidin (Cat. # PHDG1, PHDH1, PHDN1, PHDR1).  These Acti-stain phalloidins label F-actin-containing structures and fibers.  Activation of Rho family proteins can be directly quantified with either our pull-down or G-LISA activation assays.  For RhoA, use the BK036 pull-down or BK 124 G-LISA.  For Rac1, use the BK035 pull-down or BK128 G-LISA.  For Cdc42, use the BK034 pull-down or BK127 G-LISA.

     

     If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com.