Measurements of Penetration and Detoxification of PS II Herbicides in Whole Leaves by a Fluorometric Method

The effect of herbicides that inhibit the photosynthetic electron transport at the photosystem II acceptor side has been analyzed in whole plants by using a fluorometric method. The data reported indicate that the apparent variable fluorescence of the induction curve normalized to the control value provides reliable information about the penetration rate and metabolic detoxification of PS II herbicides in whole plants

Structure of the ATP-Synthase from Chloroplasts and Mitochondria Studied by Electron Microscopy

The structure of the ATP-synthase, F0F1 , from spinach chloroplasts and beef heart mitochondria has been investigated by electron microscopy with negatively stained specimens. The detergent-solubilized ATP-synthase forms string-like structures in which the F0 parts are aggregated. In most cases, the F, parts are arranged at alternating sides along the string. The F0 part has an approximate cylindrical shape with heights of 8.3 and 8.9 nm and diameters of 6.2 and 6.4 nm for the chloroplast and mitochondrial enzyme, respectively. The F, parts are disk-like structures with a diameter of about 11.5 nm and a height of about 8.5 nm. The F, parts are attached to the strings, composed of Fn parts, in most cases, with their smallest dimension parallel to the strings. The stalk connecting F0 and F, has a length of 3.7 nm and 4.3 nm and a diameter of 2.7 nm and 4.3 nm for the chloroplast and mitochondrial enzyme, respectively

Effect of Tentoxin on the Activation and on the Catalytic Reaction of Reconstituted H+-ATPase from Chloroplasts

The proton-translocating ATPase from chloroplasts, CF0F1, was isolated, purified and reconstituted into asolectin liposomes. The effect of the energy transfer inhibitor, tentoxin, on different functions of the enzyme was investigated. Tentoxin does not inhibit the nucleotide release during energization by a pH /ΔΨ jump, i.e. the activation of the enzyme is not influenced. ATP synthesis driven by a pH /ΔΨ jump and multi-site ATP hydrolysis are completely inhibited by tentoxin, whereas uni-site ATP hydrolysis is not influenced

The activity of the ATP synthase from Escherichia coli is regulated by the transmembrane proton motive force.

The ATP synthase from Escherichia coli was reconstituted into liposomes from phosphatidylcholine/phosphatidic acid. The proteoliposomes were energized by an acid-base transition and a K(+)/valinomycin diffusion potential, and one second after energization, the electrochemical proton gradient was dissipated by uncouplers, and the ATP hydrolysis measurement was started. In the presence of ADP and P(i), the initial rate of ATP hydrolysis was up to 9-fold higher with pre-energized proteoliposomes than with proteoliposomes that had not seen an electrochemical proton gradient. After dissipating the electrochemical proton gradient, the high rate of ATP hydrolysis decayed to the rate without pre-energization within about 15 s. During this decay the enzyme carried out approximately 100 turnovers. In the absence of ADP and P(i), the rate of ATP hydrolysis was already high and could not be significantly increased by pre-energization. It is concluded that ATP hydrolysis is inhibited when ADP and P(i) are bound to the enzyme and that a high Delta mu(H(+)) is required to release ADP and P(i) and to convert the enzyme into a high activity state. This high activity state is metastable and decays slowly when Delta mu(H(+)) is abolished. Thus, the proton motive force does not only supply energy for ATP synthesis but also regulates the fraction of active enzymes

Phosphatase activity of H+-ATPase from chloroplasts

AbstractThe chloroplast H+-ATPase (CF0F1) was purified from spinach chloroplasts and studied both in the soluble state and after reconstitution into asolectin liposomes. Hydrolysis of ATP and of p-nitrophenylphosphate (p-NPP) catalysed by CF0F1 was investigated. (1) Soluble, isolated CF0F1 catalyzes ATP hydrolysis and p-NPP hydrolysis. (2) ATP inhibits the phosphatase activity in the latent state (Ki=1.7 mM). (3) Addition of 100 mM sulfite increases the rate of ATP hydrolysis by a factor of 10, while p-NPP hydrolysis is completely abolished. (4) CF0F1 reconstituted into asolectin vesicles catalyzes ATP hydrolysis and p-NPP hydrolysis. When the enzyme is brought into its active state by a ΔpH/Δϕ jump, ATP hydrolysis is increased by a factor of 8, and p-NPP hydrolysis is completely abolished. (5) ATP hydrolysis by the activated enzyme is inhibited by p-NPP (Ki=1.6 mM). (6) p-NPP also inhibits ATP synthesis by the activated enzyme, competing with phosphate (Ki=0.9 mM). These results show that in the active state of CF0F1, p-NPP is not hydrolyzed but acts as a competitive inhibitor; in the inactive state of CF0F1, p-NPP is hydrolyzed. Hydrolysis of p-NPP might be used as an assay for the inactive forms of CF0F1

Serum neurofilament dynamics predicts neurodegeneration and clinical progression in presymptomatic Alzheimer's disease

Neurofilament light chain (NfL) is a promising fluid biomarker of disease progression for various cerebral proteopathies. Here we leverage the unique characteristics of the Dominantly Inherited Alzheimer Network and ultrasensitive immunoassay technology to demonstrate that NfL levels in the cerebrospinal fluid (n = 187) and serum (n = 405) are correlated with one another and are elevated at the presymptomatic stages of familial Alzheimer's disease. Longitudinal, within-person analysis of serum NfL dynamics (n = 196) confirmed this elevation and further revealed that the rate of change of serum NfL could discriminate mutation carriers from non-mutation carriers almost a decade earlier than cross-sectional absolute NfL levels (that is, 16.2 versus 6.8 years before the estimated symptom onset). Serum NfL rate of change peaked in participants converting from the presymptomatic to the symptomatic stage and was associated with cortical thinning assessed by magnetic resonance imaging, but less so with amyloid-β deposition or glucose metabolism (assessed by positron emission tomography). Serum NfL was predictive for both the rate of cortical thinning and cognitive changes assessed by the Mini-Mental State Examination and Logical Memory test. Thus, NfL dynamics in serum predict disease progression and brain neurodegeneration at the early presymptomatic stages of familial Alzheimer's disease, which supports its potential utility as a clinically useful biomarker