Phytanic acid impairs mitochondrial respiration through protonophoric action

Refsum disease is a rare, inherited neurodegenerative disorder characterized by accumulation of the dietary branched-chain fatty acid phytanic acid in plasma and tissues caused by a defect in the alphaoxidation pathway. The accumulation of phytanic acid is believed to be the main pathophysiological cause of the disease. However, the exact mechanism(s) by which phytanic acid exerts its toxicity have not been resolved. In this study, the effect of phytanic acid on mitochondrial respiration was investigated. The results show that in digitonin-permeabilized fibroblasts, phytanic acid decreases ATP synthesis, whereas substrate oxidation per se is not affected. Importantly, studies in intact fibroblasts revealed that phytanic acid decreases both the mitochondrial membrane potential and NAD(P)H autofluorescence. Taken together, the results described here show that unesterified phytanic acid exerts its toxic effect mainly through its protonophoric action, at least in human skin fibroblasts. Received 4 August 2007; received after revision 26 September 2007; accepted 10 October 2007 J. C. Komen, F. Distelmaier: These authors contributed equally to this work.     

Cellular internalization of proinsulin C-peptide

Proinsulin C-peptide is known to bind specifically to cell membranes and to exert intracellular effects, but whether it is internalized in target cells is unknown. In this study, using confocal microscopy and immunostained or rhodamine-labeled peptide, we show that C-peptide is internalized and localized to the cytosol of Swiss 3T3 and HEK-293 cells. In addition, transport into nuclei was found using the labeled peptide. The internalization was followed at 37°C for up to 1 h, and was reduced at 4°C and after preincubation with pertussis toxin. Hence, it is concluded to occur via an energy-dependent, pertussis toxin-sensitive mechanism and without detectable degradation within the experimental time course. Surface plasmon resonance measurements demonstrated binding of HEK-293 cell extract components to C-peptide, and subsequent elution of bound material revealed the components to be intracellular proteins. The identification of C-peptide cellular internalization, intracellular binding proteins, absence of rapid subsequent C-peptide degradation and apparent nuclear internalization support a maintained activity similar to that of an intracrine peptide hormone. Hence, the data suggest the possibility of one further C-peptide site of action. Received 31 October 2006; received after revision 27 December 2006; accepted 30 December 2006     

Zur Blutdruckwirkung von Aludrin und Adrenalin

Summary The depressor action of small doses of adrenaline on the blood pressure (B.P.) of the decapitated cat may be abolished by pretreatment with pilocarpine; atropine restores the original depressor action. The fall in B.P. due to adrenaline after ergotamine once more becomes an increase when pilocarpine has been given earlier. After atropine a fall in B.P. again occurs. This effect of pilocarpine and atropine on a depressor response has already been described in the case of Aludrine (N-isopropyl-noradrenaline) byFromherz.     

Über ein Chromoplastin aus reifenden Tomaten

Summary (1) By precipitation with ammonium sulfate, streptomycin, or calcium salts, we obtained from the pulp of tomatoes a substance containing carotinoids. The behaviour of this substance was analogous to that of chloroplst-substance and to that of animal cytoplasmatic nucleoproteins. Like these it contains proteins, lipids, and very probably nucleic acids. We regard this substance as achromoplastine.(2) Experiments with slices of green tomatoes show that the changing over of the chlorophyll content into the carotinoid content is inhibited by the presence of streptomycin.(3) Streptomycin inhibits the formation of chlorophyll in etiolated separated cabbage leafs, just as this drug inhibits the formation of chlorophyll in growing seeds.(4) The development of anthocyanides is not influenced by streptomycin.