PMID: 9546653Apr 18, 1998Paper

Detection of abscisic-acid-binding proteins in the microsomal protein fraction of Arabidopsis thaliana with abscisic-acid-protein conjugates used as affinity probes

European Journal of Biochemistry
J PedronE Miginiac

Abstract

A family of affinity probes has been generated to detect and purify abscisic-acid (ABA)-binding proteins, by coupling ABA onto carrier proteins (ovalbumin or BSA) through the C1 carboxyl group or the C4' carbonyl group of ABA. ELISA detection showed that these ABA-protein conjugates bound efficiently to the solubilized microsomal protein fraction of Arabidopsis thaliana, but not to the soluble protein fraction. Heat or proteolytic treatments inhibited the binding of the conjugates, indicating the protein nature of these binding sites. After membrane purification of the microsomes, the binding sites were found to be preferentially located in the plasma membrane fraction. The binding of the conjugates was independent of the nature of the carrier protein or the ABA-carrier protein linker, but was competitively inhibited with an anti-ABA mAb. Furthermore, the competitive inhibition of the binding of the conjugates with ABA, but not with the inactive ABA methyl ester analog, demonstrated the specificity of the binding and the saturability of the binding sites. The binding of the conjugates was strictly correlated to the ABA/carrier protein molar coupling ratio, confirming that the affinity of the conjugates to the ABA-binding protei...Continue Reading

Citations

Jan 1, 2002·The Arabidopsis Book·Ruth R Finkelstein, Christopher D Rock
Aug 31, 2001·Bioorganic & Medicinal Chemistry Letters·Y TodorokiN Hirai
Apr 6, 2006·The Plant Cell·Ruth R Finkelstein
Jul 15, 2015·International Journal of Molecular Sciences·Chae Woo LimSung Chul Lee
Sep 20, 2011·Plant, Cell & Environment·Sung Chul Lee, Sheng Luan
May 11, 2011·Journal of Integrative Plant Biology·Jianjun GuoJin-Gui Chen
Dec 31, 2003·The Journal of Biological Chemistry·Fawzi A RazemRobert D Hill
Mar 11, 1999·Journal of Receptor and Signal Transduction Research·M ItenE Grill
Mar 30, 1999·European Journal of Biochemistry·M BraultE Miginiac
Oct 18, 2001·Journal of Experimental Botany·D P ZhangY Y Shen
Mar 6, 1999·Current Opinion in Plant Biology·E Grill, A Himmelbach
Feb 11, 2005·Journal of the American Chemical Society·James M NyanguluSuzanne R Abrams
Dec 5, 1998·Science·E Grill, H Ziegler

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