Membrane-associated phosphoinositides-specific phospholipase C forms from Catharanthus roseus transformed roots.
We have previously reported that Catharanthus roseus transformed roots contain at least two phosphatidylinositol 4,5-bisphosphate-phospholipase C (PLC) activities, one soluble and the other membrane associated. Detergent, divalent cations, and neomycin differentially regulate these activities and pure protein is required for a greater understanding of the function and regulation of this enzyme. In this article we report a partia purification of membrane-associated PLC. We found that there are at least two forms of membraneassociated PLC in transformed roots of C. roseus. These forms were separated on the basis of their affinity for heparin. One form shows an affinity for heparin and elutes at approx 600 mM KCl. This form has a molecular mass of 67 kDa by size exclusion chromatography and Western blot analysis, whereas the other form does not bind to heparin and has a molecular mass of 57 kDa. Possible differential regulation of these forms during transformed root growth is discussed.
Echevarría-Machado
Ileana
I
Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigaciòn Científica de Yucatán A.C., Calle 43 No. 130, Chuburná de Hidalgo, C. P. 97200, Mérida, Yucatán, México.
Martínez-Estévez
Manuel
M
Muñoz-Sánchez
J Armando
JA
Loyola-Vargas
Víctor M
VM
Hernández-Sotomayor
S M Teresa
SM
De Los Santos-Briones
César
C
D015153:Blotting, Western; D030008:Catharanthus / Q000201:enzymology; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D043265:Phosphatidylinositol Diacylglycerol-Lyase / Q000302:isolation & purification / Q000378:metabolism*; D018517:Plant Roots / Q000201:enzymology*
D043265:Phosphatidylinositol Diacylglycerol-Lyase