Multiple Transport Components for Putrescine in Escherichia coli

Putrescine uptake was studied in cultures of Escherichia coli K-12 grown in media of high or low osmolarity. When grown in high osmolarity medium, a transport system of low K(m) and low V(max) was found. For cultures grown in a medium of low osmolarity, the kinetics of putrescine uptake was more complex and consistent with the existence of an additional transport system of higher K(m) and V(max). This conclusion is supported by the isolation of mutants in which one or the other system appears to be defective and by the ability of chloramphenicol to block the expression of the second transport system. Both systems appear to prefer putrescine over other compounds, since several basic amino acids and other polyamines competed only weakly for transport. The action of both uptake systems was shown to cause significant displacement of intracellular putrescine. Both systems also are at least partially energy dependent.     

Photoreceptor Spike Responses in the Hardshell Clam, Mercenaria mercenaria

Spikes were recorded from single axons in the siphonal nerve of the hardshell clam Mercenaria mercenaria which respond to dimming of light. No axons were found to respond to the onset, or increase, of illumination. In a dark-adapted state there is little or no ongoing spike activity. The responsive area of a single axon is a circle of approximately 85 µm diameter on the inner siphon wall. The number of spikes elicited at the off of constant-duration flashes grows as approximately the 0.4 power of flash intensity. For constant intensity and constant light-time fraction, the off-response increases with increasing duration at least up to 500 s duration. For long durations, the response grows as the logarithm of stimulus duration. Subthreshold light can suppress the off-response from preceding illumination. In a light-adapted state, the off-response is greater and its latency shorter than in the dark-adapted state. The fine structure of groups of cell processes thought to comprise the photoreceptor in Mercenaria is described. On the basis of morphological and physiological findings it is suggested that phototransduction occurs in the fine distal processes of the axons from which we have recorded. Axonal processes were found to contain well organized pentalaminar whorls which may be the site of photo-pigment concentration. The action spectrum obtained from the integrated responses of nerve bundles appears to be that of a single Dartnall pigment having maximal absorption at about 510 nm.     

Identification of erythro-beta-hydroxyasparagine in the EGF-like domain of human C1r

Previous studies [(1987) Biochem. J. 241, 711-720] have shown that position 150 of human C1r is occupied by a modified amino acid that, after acid hydrolysis, yields erythro-beta-hydroxyaspartic acid. In view of further investigations on the nature of this residue, peptide CN1a T8/T9 TL8 (positions 147-155) was isolated from C1r A chain by CNBr cleavage followed by enzymatic cleavages by trypsin and thermolysin. Amino acid analysis, sequential Edman degradation and FAB-MS of this peptide indicate that the residue at position 150 is an erythro-beta-hydroxyasparagine resulting from post-translational hydroxylation of asparagine.     

sn-1,2-Diacylglycerol kinase of Escherichia coli. Structural and kinetic analysis of the lipid cofactor dependence

The lipid cofactor requirement of Escherichia coli sn-1,2-diacylglycerol kinase was studied using a beta-octylglucoside mixed micellar assay (Walsh, J. P., and Bell, R. M. (1986) J. Biol. Chem. 261, 6239-6247). The enzyme was shown to have an absolute requirement for a lipid activator. sn-1,2-Dioleoylglycerol was both an activator and a substrate for the enzyme, 1,3-dioleoylglycerol was an activator but not a substrate, and sn-1,2-dioctanoylglycerol was a substrate but not an activator. Activation was observed with a large number of phospholipids, sulfolipids, neutral lipids, and detergents. Lipids with longer alkyl/acyl chains stimulated activity to a greater extent and at lower concentrations than their shorter chain homologs. Anionic lipids were the best activators, and neutral lipids were somewhat less effective. Cationic lipids were poor activators. Lipid activation was cooperative in all cases, with Hill coefficients ranging from 2.9 to 4.7. Lipid activators stabilized the enzyme against inactivation induced by diacylglycerols. The effectiveness of several lipids in stabilizing the enzyme correlated with their effectiveness as kinetic activators, suggesting a common mechanism. Kinetic analyses also suggested that a lipid cofactor-induced conformational change occurs as a part of the activation process. beta-Octylglucoside was shown not to function as a lipid cofactor for diacylglycerol kinase. The requirement for detergent in the assay was related, instead, to the need to disperse and deliver water-insoluble substrates and cofactors to the enzyme. beta-Octylglucoside also provided an inert matrix to which lipid substrates and cofactors could be added, enabling study of their concentration dependencies.     

Neuronal and Nonneuronal Contributions to Renal Catecholamine Content in the Dog

Endogenous noradrenaline and 3,4-dihydroxyphenylethylamine (dopamine) levels were measured in different zones of the dog kidney following chronic unilateral renal denervation. In outer and inner renal cortex, and in outer medulla, greater than 95% of the tissue content of both catecholamines was contributed by renal nerves, whereas in inner medulla only nonneuronal catecholamines were found. The amounts of neuronal dopamine present in outer renal cortex were greater than would be expected for a population of solely noradrenergic nerves.     

Effect of 9.6-GHz pulsed microwaves on the orb web spinning ability of the cross spider (Araneus diadematus)

Eight cross spiders (Araneus diadematus) were exposed overnight (16 h) during web-building activity to pulsed 9.6-GHz microwaves at average power densities of 10, 1, and 0.1 mW/cm2 (estimated SARs 40, 4, and 0.4 mW/g). Under these conditions, 9.6-GHz pulsed microwaves did not affect the web-spinning ability of the cross spider.