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991.
Ras and Raf-1 are key proteins involved in the transmission of developmental and proliferative signals generated by receptor and nonreceptor tyrosine kinases. Genetic and biochemical studies demonstrate that Raf-1 functions downstream of Ras in many signaling pathways. Although Raf-1 directly associates with GTP-bound Ras, an effect of this interaction on Raf-1 activity in vivo has not been established. To examine the biological consequence of the Ras/Raf-1 interaction in vivo, we set out to identify key residues of Raf-1 required for Ras binding. In this report, we show that a single amino acid mutation in Raf-1 (Arg89 to Leu) disrupted the interaction with Ras in vitro and in the yeast two-hybrid system. This mutation prevented Ras-mediated but not tyrosine kinase-mediated enzymatic activation of Raf-1 in the baculovirus/Sf9 expression system. Furthermore, kinase-defective Raf-1 proteins containing the Arg89-->Leu mutation were no longer dominant-inhibitory or capable of blocking Ras-mediated signal transduction in Xenopus laevis oocytes. These results demonstrate that the association of Raf-1 and Ras modulates both the kinase activity and the biological function of Raf-1 and identify Arg89 as a critical residue involved in this interaction. In addition, the finding that tyrosine kinases can stimulate the enzymatic activity of Raf-1 proteins containing a mutation at the Ras-interaction site suggests that Raf-1 can be activated by Ras-independent pathways. 相似文献
992.
JD Edwards PS Dovgan JM Rowley DK Agrawal PE Thorpe TE Adrian 《Canadian Metallurgical Quarterly》1994,8(6):729-734
Endothelin-1 (ET-1) is a potent vasoconstrictive polypeptide produced from vascular endothelial cells. The effects of ischaemia, reperfusion, and exsanguination on plasma ET-1 levels were studied and compared in the mongrel dog after infrarenal aortic cross clamping. Ischaemia produced a trend toward increased ET-1 serum levels (p < 0.07 with Bonferroni correction) that did not reach significance. Plasma ET-1 levels were significantly increased during reperfusion and even further elevations were found following exsanguination. We found a 2-3 fold increase in ET-1 levels following reperfusion (Initial 3.19 +/- 0.27 pg/ml vs. Reperfusion maximum 6.32 +/- 0.72 pg/ml, Bonferroni p < 0.01). Haemorrhagic shock was associated with a 3-4 fold increase in ET-1 levels (Initial 3.19 +/- 0.27 pg/ml vs. Exsanguination maximum 8.37 +/- 0.97 pg/ml Bonferroni p < 0.001). These data reveal that ET-1 is released during reperfusion and exsanguination and may mediate remote vascular events associated with infrarenal aortic cross clamping and acute blood loss. 相似文献
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NY Kiang JM Rho CC Northrop MC Liberman DK Ryugo 《Canadian Metallurgical Quarterly》1982,217(4555):175-177
A horseradish peroxidase technique was used to trace the peripheral terminations of two types of ganglion cells in adult cats. It was found that large, usually bipolar ganglion cells end on inner hair cells and small, usually pseudomonopolar ganglion cells end on outer hair cells. Thus, a virtually complete segregation of afferent neural inputs from the two types of hair cells was directly confirmed. 相似文献
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DK Rex 《Canadian Metallurgical Quarterly》1994,38(1):60-67
The dynamics of intestinal absorption, blood concentration and distribution of thiamin, biotin, nicotinate, riboflavin, pantothenate, various folates (folic acid, folinic acid, pteroyltriglutamate), vitamins A, E, C, B12, and B6 were monitored in 12 patients by multiple simultaneous sampling of blood obtained by combined catheterization of portal vein, hepatic vein, and femoral artery after vitamin ingestion. All water-soluble vitamins proved elevated after vitamin ingestion principally in portal blood within 10 minutes as compared with hepatic and femoral blood. Elevated vitamin levels in portal blood--compared to hepatic and femoral blood--remained high even after 120 min. indicating that absorption from the gut was still progressing. In contrast, ingestion of the fat-soluble vitamins A and E evoked no elevated vitamin activity in portal blood. Within 10 min. after vitamin ingestion, all folates were converted into reduced and methylated 5-methyltetrahydrofolate (5-CH3THF) on passage through the gut. At this time, portal blood elevation of 5-CH3THF persisted before its elevation in hepatic or femoral blood. Presumably, the elevation was not due to the flushing of stored 5-CH5THF from tissues but rather of folate conversion to 5-CH3THF upon gut passage. The significance of these findings is discussed. 相似文献
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