Plasmalemma from the roots of cucumber: Isolation by two-phase partitioning and characterization

1987-2
Memon, Abdul Razaque
Sommarin, Marianne
Kylin, Anders
Show full item record
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
112
views
0
downloads
Plasmalemma was isolated from the roots of 2‐week‐old cucumber plants (Cucumis sativus L. cv. Rhensk druv) by utilizing an aqueous polymer two‐phase system with 6.5%:6.5% (w/w) Dextran T500 and polyethylene glycol (PEG) 3350 at pH 7.8. The plasmalemma fraction comprised ca 6% of the membrane proteins contained in the microsomal fraction. The specific activity of the plasma membrane marker enzyme (K+, Mg2+‐ATPase) was 14‐ to 17‐times higher in the upper (PEG‐rich) than in the lower (Dextran‐rich) phase, and the reverse was true for marker enzymes (cytochrome c oxidase, EC 1.9.3.1, and antimycin A‐resistant NADPH cytochrome c reductase) of intracellular membranes. The ATPase was highly stimulated by the addition of detergent (Triton X‐100), so that the isolated plasmalemma vesicles appear tightly sealed and in a right‐side‐out orientation. Further characterization of the ATPase activities showed a pH optimum at 6.0 in the presence of Mg2+. This optimum was shifted to pH 5.8 after addition of K+. K+ stimulated the ATPase activity below pH 6 and inhibited above pH 6. The ATPase activity was specific for ATP and sensitive to N,N‐dicyclohexylcarbodiimide and sodium vanadate, with K+ enhancing the vanadate inhibition. The enzyme was insensitive to sodium molybdate, NO−3, azide and oligomycin. No Ca2+‐ATPase was detected, and even as little as 0.05 mM Ca2+ inhibited the Mg2+‐ATPase activity.
Plant Science, Genetics, Cell Biology, Physiology, General Medicine
https://hdl.handle.net/11511/52175
Physiologia Plantarum
Department of Biology, Article

Suggestions

Genetic transformation of lentil ( Lens culinaris m. cv.Sultan.1) with a transcription factor regulator (MBF1c) and analysis of transgenic plants
Kamçı, Hamdi; Çelikkol Akçay, Ufuk; Kamçı, Hamdi; Department of Biotechnology (2011)
Agrobacterium mediated genetic transformation of lentil Sultan 1 cultivar with MBF1c and evaluation of transgenic plants was aimed. The study was initially based on optimized protocol with Agrobacterium tumefaciens KYRT1 strain and pTJK136 binary plasmid. Based on this protocol and transient marker gene expression in embryo apex, 15% stable transformation efficiency was aimed. However limited knowledge about pTJK136 and problem with curing KYRT1 leaded us to use Agrobacterium tumefaciens C58C1 strain and al...
Bioproduction, structure elucidation and in vitro antiproliferative effect of eumelanin pigment from Streptomyces parvus BSB49
Bayram, Sinan; Dengiz, Çağatay; GERÇEK, Yusuf Can; ÇETİN, İdil; TOPÇUL, Mehmet Rıfkı (Springer Science and Business Media LLC, 2020-01-01)
In this study, the structure of the purified extracellular eumelanin pigment isolated from Streptomyces spp. was elucidated by detailed analysis via two different spectroscopic techniques (FT-IR and NMR). In vitro antiproliferative effects of eumelanin were evaluated on HeLa cell line. These experiments were carried out with the evaluation of the parameters including cell viability, cell index, and mitotic index. With the cell viability and cell index, IC50 concentration of eumelanin was determined as 10 μM...
Optimisation of agrobacterium mediated gene transfer and micrografting systems in lentil (lens culinaris medik)
Kamçı, Hamdi; Öktem, Hüseyin Avni; Department of Biotechnology (2004)
In this work Agrobacterium (KYRT1::pTJK136) mediated gene transfer to lentil (Lens culinaris Medik.) embriyo apex and regeneration through micro-grafting in lentil was studied. In micro-grafting two different types root stock stem height and root stock preparations were optimized. According to the results half stem length was found to be more successful then the full. Also lentil root stock was more successful then the chickpea root stock. The types of root stock preparations studied were designated as Z an...
Metal Tolerance Protein 8 Mediates Manganese Homeostasis and Iron Reallocation during Seed Development and Germination
Eroğlu, Seçkin; Meier, Bastian; Takahashi, Michiko; Terada, Yasuko; Ignatyev, Konstantin; Andresen, Elisa; Kupper, Hendrik; Peiter, Edgar; Von Wiren, Nicolaus (American Society of Plant Biologists (ASPB), 2017-07-01)
Metal accumulation in seeds is a prerequisite for germination and establishment of plants but also for micronutrient delivery to humans. To investigate metal transport processes and their interactions in seeds, we focused on METAL TOLERANCE PROTEIN8 (MTP8), a tonoplast transporter of the manganese (Mn) subclade of cation diffusion facilitators, which in Arabidopsis (Arabidopsis thaliana) is expressed in embryos of seeds. The x-ray fluorescence imaging showed that expression of MTP8 was responsible for Mn lo...
Evaluation of photosynthetic performance of wheat cultivars exposed to boron toxicity by the JIP fluorescence test
Oz, M. T.; TURAN, ÖZLEM; Kayihan, C.; EYİDOĞAN, FÜSUN; EKMEKÇİ, YASEMİN; YÜCEL, MUSTAFA; Öktem, Hüseyin Avni (Institute of Experimental Botany, 2014-12-01)
The changes in growth and photosynthetic performance of two wheat (Triticum aestivum L.) cultivars (Bolal-2973 and Atay-85) differing in their sensitivity to boron (B) toxicity were investigated under toxic B conditions. Eight-day old seedlings were exposed to highly toxic B concentrations (5, 7.5, and 10 mM H3BO3) for 5 and 9 days. Fast chlorophyll a fluorescence kinetics was determined and analysed using JIP test. Growth parameters, tissue B contents, and membrane damage were measured at two stress durati...
Citation Formats
A. R. Memon, M. Sommarin, and A. Kylin, “Plasmalemma from the roots of cucumber: Isolation by two-phase partitioning and characterization,” Physiologia Plantarum, pp. 237–243, 1987, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/52175.
METU IIS - Integrated Information Service open@metu.edu.tr