A spectrophotometric procedure is described which can be utilized to determine quantitatively chlorophyll a, chlorophyll b, pheophytin a, pheophytin b, total chlorophyll, total pheophytin, and per cent retention of chlorophylls.The method utilizes determined specific absorptivities and changes in specific absorptivity for the four components at appropriate wave lengths in 80% acetone. In addition to these specified
The use of diphenylcarbazide as an electron donor coupled to the photoreduction of 2,6-dichlorophenolindophenol by tris-washed chloroplasts or subchloroplast fragments provides a simple and sensitive assay for photosystem 2 of chloroplasts. By varying the concentration of tris buffer at pH 8.0 during an incubation period it is shown that the destruction of oxygen evolution activity is accompanied by a corresponding emergence of an ability to photooxidize diphenylcarbazide, as evidenced by absorbance changes due to diphenylcarbazide at 300 nm. The temperature-sensitive oxidation of diphenylcarbazide is inhibited by DCMU and by high ionic strengths. This activity appears to measure the primary photochemical reaction of photosystem 2.
We propose a molecular model for phospholipid membrane lysis by the ubiquitous plant toxins called thionins. Membrane lysis constitutes the first major effect exerted by these toxins that initiates a cascade of cytoplasmic events leading to cell death. X-ray crystallography, solution nuclear magnetic resonance (NMR) studies, small angle X-ray scattering and fluorescence spectroscopy provide evidence for the mechanism of membrane lysis. In the crystal structures of two thionins in the family, alpha(1)- and beta-purothionins (MW: approximately 4.8 kDa), a phosphate ion and a glycerol molecule are modeled bound to the protein. (31)P NMR experiments on the desalted toxins confirm phosphate-ion binding in solution. Evidence also comes from phospholipid partition experiments with radiolabeled toxins and with fluorescent phospholipids. This data permit a model of the phospholipid-protein complex to be built. Further, NMR experiments, one-dimensional (1D)- and two-dimensional (2D)-total correlation spectroscopy (TOCSY), carried out on the model compounds glycerol-3-phosphate (G3P) and short chain phospholipids, supported the predicted mode of phospholipid binding. The toxins' high positive charge, which renders them extremely soluble (>300 mg/mL), and the phospholipid-binding specificity suggest the toxin-membrane interaction is mediated by binding to patches of negatively charged phospholipids [phosphatidic acid (PA) or phosphatidyl serine (PS)] and their subsequent withdrawal. The formation of proteolipid complexes causes solubilization of the membrane and its lysis. The model suggests that the oligomerization may play a role in toxin's activation process and provides insight into the structural principles of protein-membrane interactions.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.