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Title: Vacuole/extravacuole distribution of soluble protease in Hippeastrum petal and Triticum leaf protoplasts

Abstract

The subcellular distribution of soluble protease in anthesis-stage, anthocyanin-containing Hippeastrum cv. Dutch Red Hybrid petal protoplasts has been reevaluated and that of Triticum aestivum L. var. Red Coat leaf protoplasts determined using /sup 125/I-fibrin as a protease substrate and improved methods for protoplast and vacuole volume estimation. Results indicate that about 20% of the Hippeastrum petal-soluble protease and about 90% of the wheat leaf-soluble protease can be assigned to the vacuole. Protoplast isolation enzyme labeled with /sup 125/I has been used to assess the efficiency of removing isolation enzyme from protoplasts by repeated washing and by separation of protoplasts from debris using density centrifugation. Results of these studies suggest that protoplasts prepared by both methods retain low levels of isolation enzyme. However, when protoplasts prepared by either method were lysed with washing medium lacking osmoticum, little isolation enzyme contaminated the lysates.

Authors:
; ;
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
OSTI Identifier:
5842916
Resource Type:
Journal Article
Journal Name:
Plant Physiol.; (United States)
Additional Journal Information:
Journal Volume: 68:5
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; PEPTIDE HYDROLASES; BIOLOGICAL LOCALIZATION; ENZYME ACTIVITY; CELL CONSTITUENTS; EXPERIMENTAL DATA; FIBRIN; IODINE 125; MEASURING METHODS; PLANT CELLS; SUBSTRATES; TRACER TECHNIQUES; ULTRACENTRIFUGATION; UPTAKE; BETA DECAY RADIOISOTOPES; CENTRIFUGATION; COAGULANTS; DATA; DAYS LIVING RADIOISOTOPES; DRUGS; ELECTRON CAPTURE RADIOISOTOPES; ENZYMES; HEMATOLOGIC AGENTS; HEMOSTATICS; HYDROLASES; INFORMATION; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; NUCLEI; NUMERICAL DATA; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; PROTEINS; RADIOISOTOPES; SCLEROPROTEINS; SEPARATION PROCESSES; 550201* - Biochemistry- Tracer Techniques; 550301 - Cytology- Tracer Techniques

Citation Formats

Wagner, G J, Mulready, P, and Cutt, J. Vacuole/extravacuole distribution of soluble protease in Hippeastrum petal and Triticum leaf protoplasts. United States: N. p., 1981. Web. doi:10.1104/pp.68.5.1081.
Wagner, G J, Mulready, P, & Cutt, J. Vacuole/extravacuole distribution of soluble protease in Hippeastrum petal and Triticum leaf protoplasts. United States. https://doi.org/10.1104/pp.68.5.1081
Wagner, G J, Mulready, P, and Cutt, J. 1981. "Vacuole/extravacuole distribution of soluble protease in Hippeastrum petal and Triticum leaf protoplasts". United States. https://doi.org/10.1104/pp.68.5.1081.
@article{osti_5842916,
title = {Vacuole/extravacuole distribution of soluble protease in Hippeastrum petal and Triticum leaf protoplasts},
author = {Wagner, G J and Mulready, P and Cutt, J},
abstractNote = {The subcellular distribution of soluble protease in anthesis-stage, anthocyanin-containing Hippeastrum cv. Dutch Red Hybrid petal protoplasts has been reevaluated and that of Triticum aestivum L. var. Red Coat leaf protoplasts determined using /sup 125/I-fibrin as a protease substrate and improved methods for protoplast and vacuole volume estimation. Results indicate that about 20% of the Hippeastrum petal-soluble protease and about 90% of the wheat leaf-soluble protease can be assigned to the vacuole. Protoplast isolation enzyme labeled with /sup 125/I has been used to assess the efficiency of removing isolation enzyme from protoplasts by repeated washing and by separation of protoplasts from debris using density centrifugation. Results of these studies suggest that protoplasts prepared by both methods retain low levels of isolation enzyme. However, when protoplasts prepared by either method were lysed with washing medium lacking osmoticum, little isolation enzyme contaminated the lysates.},
doi = {10.1104/pp.68.5.1081},
url = {https://www.osti.gov/biblio/5842916}, journal = {Plant Physiol.; (United States)},
number = ,
volume = 68:5,
place = {United States},
year = {Sun Nov 01 00:00:00 EST 1981},
month = {Sun Nov 01 00:00:00 EST 1981}
}