Tableau de caractéristiques principal
|Description||Rap1 Activation Assay Kit|
|Application||Non-radioactive Rap1 Activation Assay Kit that uses Ral GDS RBD, agarose (Catalog # 14-455) to precipitate Rap1-GTP from cell lysates & detection by a Rap1 specific antibody.|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Material Size||1 kit|
|Material Package||Kit capacity: 20-30 assays|
Rap1 Activation Assay Kit FDS
|Titre||Numéro de lot|
|Rap1 Activation Assay Kit||2918797|
|Rap1 Activation Assay Kit||3011042|
|Rap1 Activation Assay Kit||2844014|
|Rap1 Activation Assay Kit||2886123|
|Rap1 Activation Assay Kit||3113359|
|Rap1 Activation Assay Kit||3117889|
|Rap1 Activation Assay Kit - 2089003||2089003|
|Rap1 Activation Assay Kit - 2140555||2140555|
|Rap1 Activation Assay Kit - 2359951||2359951|
|Rap1 Activation Assay Kit - 2388888||2388888|
|Aperçu de la référence bibliographique||Nº PubMed|
|Measurement of GTP-bound Ras-like GTPases by activation-specific probes.|
van Triest, M, et al.
Meth. Enzymol., 333: 343-8 (2001) 2001
|Signaling of hepatocyte growth factor/scatter factor (HGF) to the small GTPase Rap1 via the large docking protein Gab1 and the adapter protein CRKL.|
Sakkab, D, et al.
J. Biol. Chem., 275: 10772-8 (2000) 2000
Afficher le résumé
Hepatocyte growth factor (HGF; scatter factor) is a multipotent protein with mitogenic, motogenic, and developmental functions. Upon activation, the HGF-receptor c-Met binds and phosphorylates the multisite docking protein Gab1. Besides binding motifs for phosphatidylinositol 3-kinase and Grb2, Gab 1 contains multiple Tyr-X-X-Pro (YXXP) motifs which, when phosphorylated, are potential binding sites for the adapter proteins c-Crk and Crk-like (CRKL). Stimulation of human embryonic kidney cells (HEK293) with HGF leads to Gab1 association with CRKL. The Gab1-CRKL interaction requires both, the SH2 domain of CRKL and the region containing the YXXP motifs in Gab1. CRKL binds via its first SH3 domain to several downstream signal transducers, including C3G an activator of the small GTPase Rap1. Indeed, Rap1 was rapidly activated after HGF stimulation of HEK293 cells. Rap1 activation through HGF was suppressed through transfection of a truncated C3G protein which only contains the SH3-binding motifs of C3G. Transfection of nonmutated Gab1 led to a strong increase of Rap1.GTP in the absence of HGF. In contrast, transfection of the GabDeltaYXXP mutant abolished the elevation of Rap1.GTP by HGF. A replating assay indicated that HGF decreases the adhesion of HEK293 cells. The results presented here delineate a novel signaling pathway from HGF to the GTPase Rap1 which depends on the interaction of the adapter protein CRKL with the exchange factor C3G and could be linked to cell migration.
|Identification of guanine nucleotide exchange factors (GEFs) for the Rap1 GTPase. Regulation of MR-GEF by M-Ras-GTP interaction|
Rebhun, J F, et al
J Biol Chem, 275:34901-8 (2000) 2000
|Rapid Ca2+-mediated activation of Rap1 in human platelets.|
Franke, B, et al.
EMBO J., 16: 252-9 (1997) 1997
Afficher le résumé
Rap1 is a small, Ras-like GTPase whose function and regulation are still largely unknown. We have developed a novel assay to monitor the active, GTP-bound form of Rap1 based on the differential affinity of Rap1GTP and Rap1GDP for the Rap binding domain of RalGDS (RBD). Stimulation of blood platelets with alpha-thrombin or other platelet activators caused a rapid and strong induction of Rap1 that associated with RBD in vitro. Binding to RBD increased from undetectable levels in resting platelets to >50% of total Rap1 within 30 s after stimulation. An increase in the intracellular Ca2+ concentration is both necessary and sufficient for Rap1 activation since it was induced by agents that increase intracellular Ca2+ and inhibited by a Ca2+-chelating agent. Neither inhibition of translocation of Rap1 to the cytoskeleton nor inhibition of platelet aggregation affected thrombin-induced activation of Rap1. In contrast, prostaglandin I2 (PGI2), a strong negative regulator of platelet function, inhibited agonist-induced as well as Ca2+-induced activation of Rap1. From our results, we conclude that Rap1 activation in platelets is an important common event in early agonist-induced signalling, and that this activation is mediated by an increased intracellular Ca2+ concentration.
|Differential interaction of the ras family GTP-binding proteins H-Ras, Rap1A, and R-Ras with the putative effector molecules Raf kinase and Ral-guanine nucleotide exchange factor|
Herrmann, C, et al
J Biol Chem, 271:6794-800 (1996) 1996
|Hallmarks of Aging|