The amino acid sequence of cytochrome c553 from Microcystis aeruginosa

Cytochrome c553 is an electron donor to P700 in the photosynthetic electron transfer chain of cyanobacteria and eukaryotic algae. We have purified this cytochrome from the cyanobacterium Microcystis aeruginosa and determined its amino acid sequence. When the amino acid sequence of this protein is compared to sequences of cytochromes c553 from other organisms, one sees that the evolution of net charge is more pronounced than the evolution of overall structure, further documenting a pronounced shift in the isoelectric point of this protein during the evolution of cyanobacteria. Cyanobacteria and algae also contain cytochrome c550 (Mr 15,500) which is quite different from cytochrome c553 (Mr 10,500). When the amino acid sequence of cytochrome c553 is compared to that of cytochrome c550, two regions of similar sequence are recognized.     

Quantitative changes in the degree of chromatin condensation during the cell cycle in differentiating Pisum sativum vascular tissue

Fixed tissue from the base of the fourth internode above the cotyledons of Pisum sativum plants were Feulgen-stained for DNA. In squash preparations three cell types: xylem vessel elements, phloem fibres and phloem sieve tube elements were identified. Measurement of Feulgen absorbance values in 0.5 X 0.5 microns measuring points across each nucleus were obtained with a scanning cytophotometer. Condensed chromatin was defined as any measuring point having an absorbance value greater than 0.32. Calculating 'percent condensed chromatin' as the percentage of the total nuclear DNA contained in those densely stained points, the pattern of change in this parameter during interphase in each cell type was observed. A generally similar pattern occurs in all three cell types. Percent condensed chromatin decreased from about 45% to 12% during G1, increased rapidly to the end of S, decreased during G2 and the following G1 to increase again during the subsequent S phase.     

Evaluation of the binding of serotonin by isolated CNS acidic lipids

Binding of serotonin by rat lipids was examined in an organic solvent-aqueous partition system. Only phospholipids and sulfatide were found to have appreciable activity: this technique was unsuitable for gangliosides due to their poor extractibility. Binding by phospholipid was abolished and that by sulfatide was greatly inhibited by increasing ionic strength in the aqueous phase. At an ionic strength of 0.3 M the apparent affinity of sulfatide for serotonin was about 3×10³ M. Both tryptamine and 5-methoxytryptamine were much more effective than serotonin in inhibiting the binding of radioactive serotonin, suggesting that the observed binding is simply a charge neutralization with little specificity. Binding of serotonin by mixed brain gangliosides was examined in an equilibrium dialysis system. Without adequate precautions, the chemical lability of serotonin was found to produce spurious data when binding was assessed by the distribution of radiolabel. Binding of serotonin by ganglioside was also greatly inhibited by increasing ionic strength: at 0.3 M an apparent affinity of about 10³ M was found. While dopamine did not inhibit the binding of radioactive serotonin, tryptamine, 5-methoxytryptamine, and serotonin were equally effective inhibitors.     

CFTR: development of high- affinity antibodies and localization in sweat gland.

Rabbit antisera were raised to six synthetic peptides corresponding to amino acid sequences contained in the protein product of the cystic fibrosis gene, CFTR. For two peptides, [Lys102]CFTR(102-116) and CFTR(1468-1480), antibody-peptide binding was of high affinity in that half-maximal binding occurred at peptide concentrations below 10 nM. Monospecific antibodies were prepared using these peptides, and these antibodies were used to stain human skin. Specific staining was detected in the cells lining the reabsorptive duct of the sweat gland. Within these lumenal cells, staining was most prominent at the apical domain but was also detected near the basolateral surface. This finding agrees well with predictions based on the effects of cystic fibrosis on sweat gland function, and suggests that these antibodies will be useful for studying CFTR in other human tissues.     

THE ISOELECTRIC POINTS OF THE PROTEINS IN CERTAIN VEGETABLE JUICES

The state in which a protein substance exists depends upon the nature of its combination with acids or bases and is changed by change in the protein compound. The nature of the compound of a protein that exists at any hydrogen ion concentration can be ascertained if the isoelectric point of the protein is known. Accordingly information regarding the isoelectric points of vegetable proteins is of importance for operations in which it may be desirable to change the state of protein substances, as in the dehydration of vegetables. The Protein in Potato Juice.—The hydrogen ion concentration of the filtered juice of the potato is in the neighborhood of 10^–7 N. Such juice contains the globulin tuberin to the extent of from 1 to 2 per cent. The character of the compound of tuberin that exists in nature was suggested by its anodic migration in an electric field. The addition of acid to potato juice dissociated this compound and liberated tuberin at its isoelectric point. The isoelectric point of tuberin coincided with a slightly lower hydrogen ion concentration than 10^–4 N. At that reaction it existed most nearly uncombined. The flow of current during cataphoresis was greatest in the neighborhood of the isoelectric point. This evidence supplements that of the direction of the migration of tuberin, since it also suggests the existence of the greatest number of uncombined ions near this point. At acidities greater than the isoelectric point tuberin combined with acid. The compound that was formed contained nearly three times as much acid as was needed to dissociate the tuberin compound that existed in nature. At such acidities tuberin migrated to the cathode. Though never completely precipitated tuberin was least soluble in the juice of the potato in the neighborhood of its isoelectric point. Both the compounds of tuberin with acids and with bases were more soluble in the juice than was uncombined tuberin. The nature of the slight precipitate that separated when potato juice was made slightly alkaline was not determined. The Protein in Carrot Juice.—The isoelectric point of the protein in carrot juice coincided with that of tuberin. Remarkably similar also were the properties of carrot juice and the juice of the potato. Existing in nature at nearly the same reaction they combined with acids and bases to nearly the same extent and showed minima in solubility at the same hydrogen ion concentrations. The greatest difference in behavior concerned the alkaline precipitate which, in the carrot, was nearly as great as the acid precipitate. The Protein in Tomato Juice.—The protein of the tomato existed in a precipitated form near its isoelectric point. Accordingly it was not present to any extent in filtered tomato juice. If, however, the considerable acidity at which the tomato exists was neutralized the protein dissolved and was filterable. It then migrated to the anode in an electric field. The addition of sufficient acid to make the hydrogen ion concentration slightly greater than 10^–5 N again precipitated the protein at its isoelectric point. At greater acidities migration was cathodic.