Vacuole/extravacuole distribution of soluble protease in Hippeastrum petal and Triticum leaf protoplasts
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.
- Research Organization:
- Brookhaven National Lab. (BNL), Upton, NY (United States)
- OSTI ID:
- 5842916
- Journal Information:
- Plant Physiol.; (United States), Vol. 68:5
- Country of Publication:
- United States
- Language:
- English
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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