scholarly journals Phosphorylation states of the (Na+ + K+)-transporting ATPase in preparations from lamb kidney and electric-eel (Electophorus electricus) electric organ

1984 ◽  
Vol 218 (2) ◽  
pp. 341-345 ◽  
Author(s):  
W E Harris ◽  
W L Stahl
Keyword(s):  
Tertiary Structure ◽  
Electric Organ ◽  
State Level ◽  
Steady State Level ◽  
Thiol Groups ◽  
Phosphorylated Form ◽  
Kidney Enzyme ◽  
Electric Eel ◽  
The Difference ◽  
Stimulation Of

Phosphorylation states of the (Na+ + K+)-transporting ATPase were studied in highly purified preparations isolated from electric-eel electric organ and from lamb kidney. The steady-state level of phosphorylated lamb kidney enzyme, obtained by reaction with [gamma-32P]ATP, was not appreciably reduced in the presence of ADP unless oligomycin was present. The phosphorylated form of the electric-eel electric-organ enzyme was reduced by at least 95% under the same conditions, suggesting that the E1P state in the kidney enzyme is more transitory than that in electric organ. The level of phosphorylation from [32P]Pi was higher in the lamb kidney preparation than in the electric-organ preparation, and the difference in stimulation of phosphorylation by ouabain in the two preparations was striking. Ouabain increased the level of phosphorylation by 35% in the kidney preparation and 734% in the electric-organ preparation. The E2P state seems to be stabilized by ouabain in the latter preparation. These findings, as well as the different reactivities of the thiol groups to blocking reagents in these preparations, suggest that the tertiary structure in the enzyme isolated from these two sources is different.

scholarly journals Modulation of Escherichia coli Adenylyl Cyclase Activity by Catalytic-Site Mutants of Protein IIAGlc of the Phosphoenolpyruvate:Sugar Phosphotransferase System

1998 ◽  
Vol 180 (3) ◽  
pp. 732-736 ◽  
Author(s):  
Prasad Reddy ◽  
Madhavi Kamireddi
Keyword(s):  
Escherichia Coli ◽  
Steady State ◽  
Adenylyl Cyclase ◽  
State Level ◽  
Cyclase Activity ◽  
Mutant Protein ◽  
Steady State Level ◽  
Adenylyl Cyclase Activity ◽  
Phosphotransferase System ◽  
Phosphorylated Form

ABSTRACT It is demonstrated here that in Escherichia coli, the phosphorylated form of the glucose-specific phosphocarrier protein IIAGlc of the phosphoenolpyruvate:sugar phosphotransferase system is an activator of adenylyl cyclase and that unphosphorylated IIAGlc has no effect on the basal activity of adenylyl cyclase. To elucidate the specific role of IIAGlcphosphorylation in the regulation of adenylyl cyclase activity, both the phosphorylatable histidine (H90) and the interactive histidine (H75) of IIAGlc were mutated by site-directed mutagenesis to glutamine and glutamate. Wild-type IIAGlc and the H75Q mutant, in which the histidine in position 75 has been replaced by glutamine, were phosphorylated by the phosphohistidine-containing phosphocarrier protein (HPr∼P) and were equally potent activators of adenylyl cyclase. Neither the H90Q nor the H90E mutant of IIAGlc was phosphorylated by HPr∼P, and both failed to activate adenylyl cyclase. Furthermore, replacement of H75 by glutamate inhibited the appearance of a steady-state level of phosphorylation of H90 of this mutant protein by HPr∼P, yet the H75E mutant of IIAGlc was a partial activator of adenylyl cyclase. The H75E H90A double mutant, which cannot be phosphorylated, did not activate adenylyl cyclase. This suggests that the H75E mutant was transiently phosphorylated by HPr∼P but the steady-state level of the phosphorylated form of the mutant protein was decreased due to the repulsive forces of the negatively charged glutamate at position 75 in the catalytic pocket. These results are discussed in the context of the proximity of H75 and H90 in the IIAGlc structure and the disposition of the negative charge in the modeled glutamate mutants.

scholarly journals Organization of thiol groups of electric-eel electric-organ sodium-plus-potassium ion-stimulated adenosine triphosphatase studied with bifunctional reagents

1980 ◽  
Vol 185 (3) ◽  
pp. 787-790 ◽  
Author(s):  
W E Harris ◽  
W L Stahl
Keyword(s):  
Adenosine Triphosphatase ◽  
Polar Solvent ◽  
Electric Organ ◽  
Thiol Group ◽  
Cross Linking ◽  
Thiol Groups ◽  
High Solubility ◽  
Efficient Inhibitor ◽  
Polar Environment ◽  
Electric Eel

The reactions of three bifunctional thiol-blocking reagents of differing cross-linking spans and two monofunctional thiol-blocking reagents with the Na+ + K+-stimulated ATPase of the electric-eel electric organ were examined. 1,5-Difluoro-2,4-dinitrobenzene with a cross-linking span of 0.3-0.5 nm (3-5 A) and high solubility in non-polar solvent was the most efficient inhibitor of enzyme activity; thus essential thiol groups exist in a non-polar environment and are approx. 0.3-0.5 nm (3-5 A) from their nearest thiol-group neighbours. Ligands promoting phosphorylation of the Na+ + K+-stimulated ATPase decreased the number of thiol groups bridged by 1,5-difluoro-2,4-dinitrobenzene and by 4,4'-difluoro-3,3'-dinitrodiphenyl sulphone [0.7-1.0 nm (7-10 A) span]. Phosphorylation is associated with a conformational change in the enzyme.

scholarly journals The third form electric organ discharge of electric eels

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jun Xu ◽  
Xiang Cui ◽  
Huiyuan Zhang
Keyword(s):  
High Voltage ◽  
Low Voltage ◽  
Electric Organ Discharge ◽  
Electric Organ ◽  
The Third ◽  
Electric Organs ◽  
Electric Eel ◽  
New Finding ◽  
Discharge Behavior ◽  
Unique Species

AbstractThe electric eel is a unique species that has evolved three electric organs. Since the 1950s, electric eels have generally been assumed to use these three organs to generate two forms of electric organ discharge (EOD): high-voltage EOD for predation and defense and low-voltage EOD for electrolocation and communication. However, why electric eels evolved three electric organs to generate two forms of EOD and how these three organs work together to generate these two forms of EOD have not been clear until now. Here, we present the third form of independent EOD of electric eels: middle-voltage EOD. We suggest that every form of EOD is generated by one electric organ independently and reveal the typical discharge order of the three electric organs. We also discuss hybrid EODs, which are combinations of these three independent EODs. This new finding indicates that the electric eel discharge behavior and physiology and the evolutionary purpose of the three electric organs are more complex than previously assumed. The purpose of the middle-voltage EOD still requires clarification.

Tacrolimus initial steady state level in post-transplant cyclophosphamide-based GvHD prophylaxis regimens

2021 ◽  
Author(s):  
Janny M. Yao ◽  
Dongyun Yang ◽  
Mary C. Clark ◽  
Salman Otoukesh ◽  
Thai Cao ◽  
...  
Keyword(s):  
Steady State ◽  
State Level ◽  
Steady State Level ◽  
Post Transplant ◽  
Gvhd Prophylaxis

Mormyromast electroreceptor organs and their afferent fibers in mormyrid fish. II. Intra-axonal recordings show initial stages of central processing

1990 ◽  
Vol 63 (2) ◽  
pp. 303-318 ◽  
Author(s):  
C. C. Bell
Keyword(s):  
Lateral Line ◽  
Electric Organ Discharge ◽  
Electric Organ ◽  
Direct Stimulation ◽  
Postsynaptic Potentials ◽  
Central Processing ◽  
Refractory Periods ◽  
Synaptic Potentials ◽  
Afferent Fibers ◽  
Stimulation Of

1. Physiologically and morphologically identified primary afferent fibers from mormyromast electroreceptor organs were recorded intracellularly. The fiber recordings were made from the nerve root of the posterior lateral line nerve, where the fibers enter the brain, and from the electrosensory lateral line lobe (ELL), near the central terminals of the fibers. 2. The intracellular recordings reveal a variety of potentials, synaptic and nonsynaptic, in addition to the large orthodromic action potentials from the periphery. The goal of the present study was to describe and interpret these various potentials in mormyromast afferent fibers as a first step in understanding the processing of electrosensory information in ELL. 3. Three types of synaptic potentials were recorded inside mormyromast afferent fibers: 1) electric organ corollary discharge (EOCD) excitatory postsynaptic potentials (EPSPs), driven by the motor command that elicits the electric organ discharge (EOD); 2) EPSPs evoked by electrosensory stimulation of electroreceptors in the skin near the electroreceptor from which the recorded fiber originates or by direct stimulation of an electrosensory nerve; and 3) inhibitory postsynaptic potentials (IPSPs) evoked by electrosensory stimulation of more distant electroreceptors. These synaptic potentials can be attributed to synaptic input to postsynaptic cells in ELL that is observed inside the afferent fibers because of electrical synapses between the fibers and the postsynaptic cells. 4. The peripherally evoked EPSPs could frequently be shown to be unitary. The unitary EPSPs were identical to the orthodromic spikes in originating from a single electroreceptor, in threshold, and in latency shift with increasing stimulus intensity. These similarities suggest that the unitary EPSPs are electrotonic EPSPs caused by impulses in other mormyromast afferent fibers that terminate on some of the same postsynaptic cells as the recorded fiber. The peripherally evoked IPSPs had a longer latency than the EPSPs or orthodromic spikes, requiring the presence of an inhibitory interneuron. 5. The peripherally evoked EPSPs, both unitary and nonunitary, show absolute refractory periods of 3-8 ms, followed by relative refractory periods of approximately 8 ms, when tested with two identical stimuli to a nerve. These refractory periods are interpreted as because of refractoriness in the fine preterminal branches of the axonal arbor. 6. A depolarizing afterpotential is commonly associated with the orthodromic spike and probably results from the successful propagation of the spike into the entire terminal arbor. The depolarizing afterpotential has a refractory period that is similar to that of the peripherally evoked EPSPs and that is also interpreted as refractoriness in the fine preterminal branches.(ABSTRACT TRUNCATED AT 400 WORDS)

Enhanced coupling of adenylate cyclase to lipolysis in permeabilized adipocytes from trained rats

1991 ◽  
Vol 71 (1) ◽  
pp. 23-29 ◽  
Author(s):  
T. Izawa ◽  
T. Komabayashi ◽  
T. Mochizuki ◽  
K. Suda ◽  
M. Tsuboi
Keyword(s):  
Adenylate Cyclase ◽  
Dose Response ◽  
Response Curve ◽  
Positive Relationship ◽  
Regression Coefficient ◽  
Permeabilized Cells ◽  
Major Mechanism ◽  
The Difference ◽  
And Control ◽  
Stimulation Of

Digitonin-permeabilized adipocytes were used to study the coupling of adenylate cyclase (AC) to lipolysis in exercise-trained rats. Isoproterenol-(IPR) stimulated lipolysis in permeabilized cells was significantly greater in trained than in control rats. Under essentially identical conditions, the dose-response curve for IPR stimulation of AC activity in the absence of 3-isobutyl-1-methylxanthine was similar in trained and control rats. However, the potency of stimulation by IPR as a percentage of the basal level was greater in trained rats. AC activity and lipolysis in the presence of 3-isobutyl-1-methylxanthine were also significantly greater in trained than in control rats. Least-squares analysis by plotting the log AC vs. lipolysis values showed that the regression coefficient was about three-fold greater in trained than in control rats. The concentration of endogenous adenosine 3′,5′-cyclic monophosphate (cAMP) needed to produce a half-maximal lipolytic response was 18.58 and 10.81 pmol.min-1.10(6) cells-1 in control and trained rats, respectively. Thus a positive relationship existed between lipolysis and AC activity, with a tighter coupling in trained rats. Lipolysis in response to exogenous cAMP tended to be greater in trained than in control rats, and the difference was statistically significant for 50 microM and 10 mM cAMP. Our finding support the concept that the major mechanism of enhanced lipolysis in trained rats was an increase in the activity of enzymatic step(s) distal to cAMP.

Responses of Medullary Dorsal Horn Neurons to Corneal Stimulation by CO2 Pulses in the Rat

1999 ◽  
Vol 82 (5) ◽  
pp. 2092-2107 ◽  
Author(s):  
Harumitsu Hirata ◽  
James W. Hu ◽  
David A. Bereiter
Keyword(s):  
Thalamic Nucleus ◽  
Receptive Fields ◽  
Male Rats ◽  
Cervical Cord ◽  
Type I ◽  
Type Ii ◽  
Upper Cervical ◽  
The Difference ◽  
Evoked Activity ◽  
Stimulation Of

Corneal-responsive neurons were recorded extracellularly in two regions of the spinal trigeminal nucleus, subnucleus interpolaris/caudalis (Vi/Vc) and subnucleus caudalis/upper cervical cord (Vc/C1) transition regions, from methohexital-anesthetized male rats. Thirty-nine Vi/Vc and 26 Vc/C1 neurons that responded to mechanical and electrical stimulation of the cornea were examined for convergent cutaneous receptive fields, responses to natural stimulation of the corneal surface by CO2 pulses (0, 30, 60, 80, and 95%), effects of morphine, and projections to the contralateral thalamus. Forty-six percent of mechanically sensitive Vi/Vc neurons and 58% of Vc/C1 neurons were excited by CO2 stimulation. The evoked activity of most cells occurred at 60% CO2 after a delay of 7–22 s. At the Vi/Vc transition three response patterns were seen. Type I cells ( n = 11) displayed an increase in activity with increasing CO2 concentration. Type II cells ( n = 7) displayed a biphasic response, an initial inhibition followed by excitation in which the magnitude of the excitatory phase was dependent on CO2 concentration. A third category of Vi/Vc cells (type III, n = 3) responded to CO2 pulses only after morphine administration (>1.0 mg/kg). At the Vc/C1 transition, all CO2-responsive cells ( n = 15) displayed an increase in firing rates with greater CO2 concentration, similar to the pattern of type I Vi/Vc cells. Comparisons of the effects of CO2 pulses on Vi/Vc type I units, Vi/Vc type II units, and Vc/C1 corneal units revealed no significant differences in threshold intensity, stimulus encoding, or latency to sustained firing. Morphine (0.5–3.5 mg/kg iv) enhanced the CO2-evoked activity of 50% of Vi/Vc neurons tested, whereas all Vc/C1 cells were inhibited in a dose-dependent, naloxone-reversible manner. Stimulation of the contralateral posterior thalamic nucleus antidromically activated 37% of Vc/C1 corneal units; however, no effective sites were found within the ventral posteromedial thalamic nucleus or nucleus submedius. None of the Vi/Vc corneal units tested were antidromically activated from sites within these thalamic regions. Corneal-responsive neurons in the Vi/Vc and Vc/C1 regions likely serve different functions in ocular nociception, a conclusion reflected more by the difference in sensitivity to analgesic drugs and efferent projection targets than by the CO2 stimulus intensity encoding functions. Collectively, the properties of Vc/C1 corneal neurons were consistent with a role in the sensory-discriminative aspects of ocular pain due to chemical irritation. The unique and heterogeneous properties of Vi/Vc corneal neurons suggested involvement in more specialized ocular functions such as reflex control of tear formation or eye blinks or recruitment of antinociceptive control pathways.

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