Three RNases in senescent and nonsenescent wheat leaves. Characterization by activity staining in sodium dodecyl sulfate-polyacrylamide gels

We have described three RNases in wheat leaves (Triticum aestivum L. cv Chinese Spring) and developed assays for measuring each RNase individually in crude leaf extracts. We initially used activity staining in sodium dodecyl sulfate-polyacrylamide gels to characterize RNases in extracts of primary a...

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Bibliographic Details
Published in:Plant physiology (Bethesda) 1991-12, Vol.97 (4), p.1402-1408
Main Authors: Blank, A. (University of Washington, Seattle, WA), McKeon, T.A
Format: Article
Language:English
Subjects:
Acetates
ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
Analytical, structural and metabolic biochemistry
Biological and medical sciences
COMPOSICION QUIMICA
COMPOSITION CHIMIQUE
Electrophoresis
ENVEJECIMIENTO
Enzymes
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Gels
Hydrolases
Imidazoles
Leaves
Metabolism
Plant physiology and development
RIBONUCLEASAS
RIBONUCLEASE
RNA
Seedlings
Sodium
TRITICUM AESTIVUM
VIEILLISSEMENT
Wheat
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Summary:We have described three RNases in wheat leaves (Triticum aestivum L. cv Chinese Spring) and developed assays for measuring each RNase individually in crude leaf extracts. We initially used activity staining in sodium dodecyl sulfate-polyacrylamide gels to characterize RNases in extracts of primary and flag leaves. We thus identified acid RNase (EC 3.1.27.1, here designated RNase WL(A)), and two apparently novel enzymes, designated RNases WL(B) and WL(C). RNase WL(B) activity displays a distinctive isozyme pattern, a molecular mass of 26 kilodaltons (major species), a broad pH range with an optimum near neutrality, insensitivity to EDTA, and stimulation by moderate concentrations of KCl and by MgCl2. RNase WL(C) activity exhibits a molecular mass of 27 kilodaltons, a neutral pH optimum, insensitivity to EDTA, and inhibition by KCl, MgCl2, and tri-(hydroxymethyl)aminomethane. Based on distinctive catalytic properties established in gels, we designed conventional solution assays for selective quantitation of each RNase activity. We used the assays to monitor the individual RNases after gel filtration chromatography and native gel electrophoresis of extracts. In accompanying work, we used the assays to monitor RNases WL(A), WL(B), and WL(C), which are present in senescent and nonsenescent leaves, during the course of leaf senescence
ISSN:0032-0889
1532-2548
DOI:10.1104/pp.97.4.1402