Arachidonoyl transacylase in human platelets. Coenzyme A-independent transfer of arachidonate from phosphatidylcholine to lysoplasmenylethanolamine

Human platelets contain an enzyme that catalyzes CoA-independent release of arachidonic acid from phosphatidylcholine with concomitant incorporation into plasmenylethanolamine. Addition of lysoplasmenylethanolamine (10-80 microM) to a crude membrane preparation of prelabeled platelets (0.24 mg of protein/ml) induces transfer of [3H]arachidonate from endogenous phosphatidylcholine to lysoplasmenylethanolamine (0.8 nmol of arachidonic acid/min/mg of protein). The transacylation reaction occurs in the absence of Ca2+, has a broad pH optimum from 7 to 8, is not affected by excess unlabeled arachidonic acid, and is inhibited by N-ethylmaleimide (0.2 mM) and Triton X-100 (0.1 mg/ml). The enzyme shows a high specificity toward the acyl donor (phosphatidylcholine), transfers fatty acids in the order: arachidonic greater than eicosatrienoic greater than oleic, and preferentially acylates lysoplasmenylethanolamine but also other lysophosphatides (lysophosphatidylethanolamine greater than lysophosphatidylserine greater than lysophosphatidylinositol = 0). Platelet acyltransferase, on the other hand, acylates ethanolamine lysophosphatides with free arachidonic acid in the order: lysophosphatidyl-ethanolamine greater than lysoplasmenylethanolamine. These results suggest that a distinct acylation mechanism exists for introduction of arachidonic acid into plasmalogen phosphatides. In stimulated platelets, the transacylase may play an additional role in the controlled release of esterified arachidonic acid for synthesis of the biologically active oxygenated metabolites.     

Comparative analysis of the 5''-end regions of two repressible acid phosphatase genes in Saccharomyces cerevisiae.

The nucleotide sequence of 5'-noncoding and N-terminal coding regions of two coordinately regulated, repressible acid phosphatase genes from Saccharomyces cerevisiae were determined. These unlinked genes encode different, but structurally related polypeptides of molecular weights 60,000 and 56,000. The DNA sequences of their 5'-flanking regions show stretches of extensive homology upstream of, and surrounding, a "TATA" sequence and in a region in which heterogeneous 5' ends of the p60 mRNA were mapped. The predicted amino acid sequences encoded by the N-terminal regions of both genes were confirmed by determination of the amino acid sequence of the native exocellular acid phosphatase and the partial sequence of the presecretory polypeptide synthesized in a cell-free protein synthesizing system. The N-terminal region of the p60 polypeptide was shown to be characterized by a hydrophobic 17-amino acid signal polypeptide which is absent in the native exocellular protein and thought to be necessary for acid phosphatase secretion.     

COMPARISON OF THE BEHAVIOUR OF A SOLUBLE AND A MEMBRANE-BOUND ENZYME IN TRANSECTED PERIPHERAL NERVES

Phosphoglucoisomerase (PGI), a soluble enzyme, and AChE, a membrane-bound enzyme were studied in transected peroneal nerves of dog and in isolated segments of these nerves. Although activities of both enzymes increased at the ends of transected nerves, marked differences in their behaviour were observed. The increment in AChE activity was much sharper than that of PGI and continued to grow with time whereas the increase in PGI developed fully within the initial hours after transection and did not change thereafter. In an isolated nerve segment AChE accumulated at both ends with a concomitant decrease in the middle part, whereas changes in PGI activity appeared only in the terminal parts, the rest of the nerve remaining at the normal level. The terminal increase of PGI did not, contrary to that of AChE, depend on the length of the isolated segment. The changes in PGI activity may be features of a local peritraumatic reaction whereas those of AChE indicate involvement of the whole segment along which the enzyme containing organelles are transported.     

Comparison of the dye method with the thermocouple psychrometer for measuring leaf water potentials

The dye method for measuring water potential was examined and compared with the thermocouple psychrometer method in order to evaluate its usefulness for measuring leaf water potentials of forest trees and common laboratory plants. Psychrometer measurements are assumed to represent the true leaf water potentials. Because of the contamination of test solutions by cell sap and leaf surface residues, dye method values of most species varied about 1 to 5 bars from psychrometer values over the leaf water potential range of 0 to −30 bars. The dye method is useful for measuring changes and relative values in leaf potential. Because of species differences in the relationships of dye method values to true leaf water potentials, dye method values should be interpreted with caution when comparing different species or the same species growing in widely different environments. Despite its limitations the dye method has a usefulness to many workers because it is simple, requires no elaborate equipment, and can be used in both the laboratory and field.     

Plasmaeinschlüsse bei einigen Desmidiaceen-Gattungen

Ohne Zusammenfassung Mit 12 Textabbildungen     

Effect of Paper on the Performance Assay in the Control of Antibiotic Sensitivity Discs

Antibiotic discs were prepared, using several several batches of papers meeting Food and Drug Administration specifications. The analysis of 1,152 zones of inhibition produced showed no performance differences among these batches. Other discs were prepared using papers of different grades. These produced large differences in performance. It is obvious, therefore, that the use of a specified disc paper is necessary for standardizing the performances of the products of various manufacturers and that reproducible results can be attained with the grade of paper specified.     

3-Diazirine-derivatives of bile salts for photoaffinity labeling

New carbene-generating photolabile bile salt derivatives, 3,3-azo-7 alpha,12 alpha-dihydroxy-5 beta [7 beta-3H]cholan-24-oic acid and (3,3-azo-7 alpha,12 alpha-dihydroxy-5 beta [7 beta-3H]cholan-24-oyl)-2- aminoethanesulfonic acid were synthesized with high specific radioactivity. These 3-diazirine-derivatives could be activated to the corresponding carbenes by irradiation with ultraviolet light at 350 nm with a half-life time of 2 min. The 3-diazirine derivatives behaved in enterohepatic circulation like the natural bile salts. The uptake of [3H]taurocholate into isolated hepatocytes was competitively inhibited by (3,3-azo-7 alpha,12 alpha-dihydroxy-5 beta-cholan-24-oyl)-2- aminoethanesulfonic acid indicating that the 3,3-azo-derivative of taurocholate shares the hepatic transport systems for natural bile salts. It was demonstrated that the radioactively labeled 3-diazirine bile salt derivatives are useful probes for photoaffinity labeling of bile salt binding proteins especially in intact cells and tissues.     

Sequence comparisons of developmentally regulated collagen genes of Caenorhabditis elegans

Collagen genes col-6, col-7 (partial), col-8, col-14 and col-19 from the nematode Caenorhabditis elegans were sequenced, and compared to the previously sequenced genes col-1 and col-2. The genes are between 1.0 and 1.2 kb in length, and each includes one or two short introns. The presumptive promoter regions contain sequences similar to the eukaryotic TATA promoter element. Two distinct, conserved sequences were found in the presumptive promoter regions of, respectively, the dauer larva-specific genes col-2 and col-6, and the primarily adult-specific genes col-7 and col-19. The domain structures of the collagen polypeptides are similar: each polypeptide contains two triple-helix forming (Gly-X-Y)n domains, one of 30-33 amino acids (aa), and the other of 127-132 aa. The latter domain is interrupted by one to three short (2-8 aa) non-(Gly-X-Y)n segments that occur at relatively conserved locations in each polypeptide. Sets of cysteine residues flank the (Gly-X-Y)n domains in all of the polypeptides. The genes can be placed into three families based upon amino acid sequence similarities. Genes within a family do not always exhibit similar developmental expression programs, suggesting that structural and regulatory regions of the genes have evolved separately. The codon usage in the genes is highly asymmetrical, with adenine appearing in the third position of 85% of the glycine codons, and 93% of the proline codons.