Processing of V-ATPase Subunit B of Mesembryanthemum crystallinum L. Is Mediated in Vitro by a Protease and/or Reactive Oxygen Species
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Rudolf Krisch
Abstract
Soluble proteins were isolated from leaves of the common ice plant Mesembryanthemum crystallinum L. in the CAM state of photosynthesis and tested for protease activity using amino acidβnaphthylamide (NA)derivatives in a search for proteolytic activity responsible for cleavage of the VATPase subunit B. This cleavage is suggested to occur at the peptide bond between Met192 and Glu193. At neutral pH MetNA was one of seven derivatives which were cleaved by proteases present in this fraction. Enzymes exhibiting proteolytic activity were separated from other soluble proteins by Superose 12-size exclusion FPLC. Incubation of partially purified protease with tonoplastenriched membrane vesicle fractions isolated from M. crystallinum in the C[3]state of photosynthesis led to a decrease in subunit B (55 kDa) protein amount and to the formation of the polypeptide D (32 kDa), which has been previously suggested to represent a fragment of subunit B. Cleavage of subunit B and the appearance of D also occurred during incubation of tonoplast vesicles in the presence of reactive oxygen species. In addition to D, the polypeptide E (28 kDa) appeared after incubation with protease and/or reactive oxygen species. Taken into account that D and E crossreacted with an affinity purified antiserum directed against subunit B, D as well as E might represent fragments of subunit B. These results open new perspectives with respect to the regulation of VATPase modification and turnover.
Copyright © 2000 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- Characterization of Regulatory Elements in the 5'-Flanking Region of the GM2 Activator Gene
- Transcriptional Activity of GLI1 Is Negatively Regulated by Protein Kinase A
- Peptide Bond Synthesis: Function of the efp Gene Product
- Molecular Cloning, Expression and Purification of Muscle Fructose-1,6-Bisphosphatase from Zaocys dhumnades: the Role of the N-Terminal Sequence in AMP Activation at Alkaline pH
- The pH-Dependent Interaction of Cinnamomin with Lipid Membranes Investigated by Fluorescence Methods
- Nitric Oxide Detection and Visualization in Biological Systems. Applications of the FNOCT Method
- Processing of V-ATPase Subunit B of Mesembryanthemum crystallinum L. Is Mediated in Vitro by a Protease and/or Reactive Oxygen Species
- The Thioredoxin Boxes of Thyroglobulin: Possible Implications for Intermolecular Disulfide Bond Formation in the Follicle Lumen
- New Bivalent Thrombin Inhibitors with N? (Methyl)Arginine at the P1-Position
- Multiple Promoters Direct the Tissue-Specific Expression of Rat Mitochondrial Glycerol-3-Phosphate Dehydrogenase
- Characterization of SNP, a Novel Tissue- and Phase-Specific Nuclear Protein Expressed during the Proliferative Phase in the Oviduct of the Lizard Podarcis sicula Raf.
- Association of Betaine-Homocysteine S-Methyltransferase with Microtubules
- Re-Evaluation of Amino Acid Sequence and Structural Consensus Rules for Cysteine-Nitric Oxide Reactivity
- Expression of Human Tissue Kallikrein in Differentiated HL-60 Cells
- Erratum
