Temperature dependence of the conductivity of individual potential-dependent K+-channels in mollusk neurons]

Using the patch-clamp method temperature dependences of the chord conductance of single potential--dependent slow and fast K+ channels in mollusk neurons were studied. Under control conditions (20 degrees C, 0 mV, [K+]o = 1.5 mM and [K+]i = 100 mM) the conductances of the fast and slow K+ channels were equal to 20-25 pS and 30-40 pS, respectively. Besides, the temperature dependences of the currents through the K+ channels of lesser conductance (5-20 pS) were studied. Some of these channels may be regarded as subtypes of the fast and slow K+ channels named above. It was found that for the channels of all types single channel currents arise with temperature. However, in the range of 10-20 degrees C an anomalous conductance decrease at temperature elevation was observed. For all channels except for the fast one at temperatures above 20 degrees C activation energy (delta Ea) calculated from the Arrhenius plots of the currents was about 4 kcal/mol. At the temperatures below 10 degrees C delta Ea was equal to about 12 kcal/mol. In this temperature range delta Ea had a pronounced potential dependency. Temperature dependences of the fast K+ channel conductance were opposite to those of the slow K+ channel to some extent.     

Membrane-Delimited Phosphorylation Enables the Activation of the Outward-Rectifying K Channels in Motor Cell Protoplasts of Samanea saman

Outward-rectifying K channels activated by membrane depolarization (Kout or KD channels) control K+ efflux from plant cells. To find out to what extent phosphorylation is required for the activity of these channels, the patch-clamp method was applied to protoplasts from the legume Samanea saman in both whole-cell and isolated-patch configurations. In the absence of either Mg2+ or ATP in the "cytosolic" solution, the KD channel activity declined completely within 15 min. This decline could be reversed in excised, inside-out patches by restoring MgATP (1 mM) to the cytoplasmic side of the membrane. Mg2+ (1 mM) plus 5[prime]-adenylylimidodiphosphate (1 mM), a nonhydrolyzable ATP analog, did not substitute for ATP. Mg2+ (1 mM) plus adenosine 5[prime]-O-(3-thiotriphosphate) (25 to <100 [mu]M), an irreversibly thiophosphorylating ATP analog, sustained channel activity irreversibly. 1-(5-IsoquinolinesulphonyI)-2- methylpiperazine (100 [mu]M), a broad-range kinase inhibitor, blocked the activity of KD channels in the presence of MgATP. These results strongly suggest that the activation of the outward-rectifying K channels by depolarization depends critically on phosphorylation by a kinase tightly associated with the KD channel.     

Reconstitution of the voltage-sensitive sodium channel of rat brain from solubilized components

The voltage-sensitive sodium channel of rat brain synaptosomes was solubilized with sodium cholate. The solubilized sodium channel migrated on a sucrose density gradient with an apparent S20,w of approximately 12 S, retained [3H]saxitoxin ([3H]STX) binding activity that was labile at 36 degrees C but no longer bound 125I-labeled scorpion toxin (125I-ScTX). Following reconstitution into phosphatidylcholine vesicles, the channel regained 125I-ScTX binding and thermal stability of [3H]STX binding. Approximately 50% of the [3H]STX binding activity and 58% of 125I-ScTX binding activity were recovered after reconstitution. The reconstituted sodium channel bound STX and ScTX with KD values of 5 and 10 nM, respectively. Under depolarized conditions, veratridine enhanced the binding of 125I-ScTX with a K0.5 of 20 microM. These KD and K0.5 values are similar to those of the native synaptosome sodium channel. 125I-ScTX binding to the reconstituted sodium channel, as with the native channel, was voltage dependent. The KD for 125I-ScTX increased with depolarization. This voltage dependence was used to demonstrate that the reconstituted channel transports Na+. Activation of sodium channels by veratridine under conditions expected to cause hyperpolarization of the reconstituted vesicles increased 125I-ScTX binding 3-fold. This increased binding was blocked by STX with K0.5 = 5 nM. These data indicate that reconstituted sodium channels can transport Na+ and hyperpolarize the reconstituted vesicles. Thus, incorporation of solubilized synaptosomal sodium channels into phosphatidylcholine vesicles results in recovery of toxin binding and action at each of the three neurotoxin receptor sites and restoration of Na+ transport by the reconstituted channels.     

Transition temperatures of the electrical activity of ion channels in the nerve membrane

The temperature dependence of some of the electrical characteristics of neuronal membranes from Aplysia giant neurons and crustacean and cuttlefish giant axons has been analyzed. Arrhenius plots for the maximum rate of depolarization of (V+max) or repolarization (V-max) of the action potential, for the resting membrane conductance, and for the speed of propagation of the action potential, exhibited clear breaks at characteristic temperatures between 17 and 20 degrees C. Lobster giant axons and frog nodes of Ranvier were voltage-clamped at different temperatures between 5 and 30 degrees C. Arrhenius plots for relaxation times related to the opening and closing processes affecting the Na+ and K+ channels were linear. No 'transition' temperature was detected. However, clear-cut changes in (Formula: see text) Na+ and K+ currents, were consistantly observed around 18 degrees C. Values for (Formula: see text) plateaued above 18 degrees C, then decreased gradually as a function of reduced temperature. Variations in temperature between 1 and 30 degrees C did not alter the binding properties of [3H]tetrodotoxin to a purified crab axonal membrane. Pharmacological properties of the Na+ channel are sensitive to temperature. The temperature-dependent effect of veratridine has been studied and indicates a change in properties of the Na+ channel below 20 degrees C. These results support the possibility that the fluidity of membrane lipids in the ionic channel microenvironment may influence the degree to which the channel can open.     

Patch-clamp studies in human macrophages: Single-channel and whole-cell characterization of two K+ conductances

Summary Human peripheral blood monocytes cultured for varying periods of time were studied using whole-cell and single-channel patch-clamp recording techniques. Whole-cell recordings revealed both an outward K current activating at potentials >20 mV and an inwardly rectifying K current present at potentials negative to –60 mV. Tail currents elicited by voltage steps that activated outward current reversed nearE _K, indicating that the outward current was due to a K conductance. TheI–V curve for the macroscopic outward current was similar to the mean single-channelI–V curve for the large conductance (240 pS in symmetrical K) calcium-activated K channel present in these cells. TEA and charybdotoxin blocked the whole-cell outward current and the single-channel current. Excised and cell-attached single-channel data showed that calcium-activated K channels were absent in freshly isolated monocytes but were present in >85% of patches from macrophages cultured for >7 days. Only 35% of the human macrophages cultured for >7 days exhibited whole-cell inward currents. The inward current was blocked by external barium and increased when [K] _o increased. Inward-rectifying single-channel currents with a conductance of 28 pS were present in cells exhibiting inward whole-cell currents. These single-channel currents are similar to those described in detail in J774.1 cells (L.C. McKinney & E.K. Gallin,J. Membrane Biol. 103:41–53, 1988).     

Temperature dependence of K(+)-channel properties in human T lymphocytes.

We have used whole-cell patch clamp to determine the temperature dependence of the conductance and gating kinetics of the voltage-gated potassium channel in quiescent, human peripheral blood T lymphocytes. Threshold for activation, steady-state inactivation, and the reversal potential are the same at 22 degrees and 37 degrees C. However, the time-constants for activation, inactivation, deactivation, and release from inactivation are quite sensitive to temperature, changing by at least a factor of five in each case over this range of temperatures. The onset of cumulative inactivation at 22 degrees and 37 degrees C reflects the time-course of deactivation. Peak outward current is approximately twofold greater at 37 degrees C than at 22 degrees C; this increase is also manifest at the single channel level. Energies of activation for conductance, activation, inactivation, deactivation, and release from inactivation are 8.2, 22.1, 25.0, 36.2, and 42.2 kcal/mol, respectively. No new channels were observed at 37 degrees C, and there was no evidence for alteration of the K+ conductance by putative modulators at 22 or 37 degrees C.     

Protein Synthesis and Breakdown during Heat Shock of Cultured Pear (Pyrus communis L.) Cells

Cultured pear (Pyrus communis L. cv Passe Crassane) cells were subjected to temperatures of 39, 42, and 45[deg]C. Heat-shock protein (hsp) synthesis was greater at 30[deg]C than at temperatures above 40[deg]C and continued for up to 8 h. Both cellular uptake of radiolabeled methionine and total protein synthesis were progressively lower as the temperature was increased. Polysome levels decreased immediately when cells were placed at 39 or 42[deg]C, although at 39[deg]C the levels began to recover after 1 h. In cells from both temperatures, reassembly occurred after transfer of cells to 25[deg]C Four heat-shock-related mRNAs[mdash]hsp17, hsp70, and those of two ubiquitin genes[mdash]all showed greatest abundance at 39[deg]C and decreased at higher temperatures. Protein degradation increased with time at 42 and 45[deg]C, but at 39[deg]C it increased for the first 2 h and then decreased. In the presence of cycloheximide, which prevented hsp synthesis, protein degradation at 39[deg]C was as great as that at 45[deg]C in the absence of cycloheximide. The data suggest that hsps may have a role in protecting proteins from degradation at the permissive temperature of 39[deg]C. At temperatures high enough to inhibit hsp synthesis, protein degradation was enhanced. Although ubiquitin may play a role in specific protein degradation, it does not appear to be involved in increased protein degradation occurring above 40[deg]C.     

Ammonia Flux between Oilseed Rape Plants and the Atmosphere in Response to Changes in Leaf Temperature,Light Intensity,and Air Humidity (Interactions with Leaf Conductance and Apoplastic NH4+ and H+ Concentrations)

NH3 exchange between oilseed rape (Brassica napus) plants and the atmosphere was examined at realistic ambient NH3 levels under controlled environmental conditions. Different leaf conductances to NH3 diffusion were obtained by changing leaf temperature (10 to 40[deg]C), light intensity (0 to 600 [mu]mol m-2 s-1), and air humidity (20 to 80%), respectively. NH3 adsorption to the cuticle with subsequent NH3 transport through the epidermis had no significant effect on the uptake of atmospheric NH3, even at 80% relative air humidity. NH3 fluxes increased linearly with leaf conductance when light intensities were increased from 0 to 600 [mu]mol m-2 s-1. Increasing leaf temperatures from 10 to 35[deg]C caused an exponential increase in NH3 emission from plants exposed to low ambient NH3 concentrations, indicating that leaf conductance was not the only factor responding to the temperature increase. The exponential relationship between NH3 emission and temperature was closely matched by the temperature dependence of the mole fraction of gaseous NH3 above the leaf apoplast (NH3 compensation point), as calculated on the basis of NH4+ and H+ concentrations in the leaf apoplast at the different leaf temperatures. NH3 fumigation experiments showed that an increase in leaf temperature may cause a plant to switch from being a strong sink for atmospheric NH3 to being a significant NH3 source. In addition to leaf temperature, the size of the NH3 compensation point depended on plant N status and was related to plant ontogeny.     

External pH effects on the depolarization-activated K channels in guard cell protoplasts of Vicia faba

Previous studies reveal that the pH of the apoplastic solution in the guard cell walls may vary between 7.2 and 5.1 in closed and open stomata, respectively. During these aperture and pH changes, massive K+ fluxes cross the cellular plasma membrane driving the osmotic turgor and volume changes of guard cells. Therefore, we examined the effect of extracellular pH on the depolarization-activated K channels (KD channels), which constitute the K+ efflux pathway, in the plasma membrane of Vicia faba guard cell protoplasts. We used patch clamp, both in whole cells as well as in excised outside-out membrane patches. Approximately 500 KD channels, at least, could be activated by depolarization in one protoplast (density: approximately 0.6 micron-2). Acidification from ph 8.1 to 4.4 decreased markedly the whole-cell conductance, GK, of the KD channels, shifted its voltage dependence, GK- EM, to the right on the voltage axis, slowed the rate of activation and increased the rate of deactivation, whereas the single channel conductance was not affected significantly. Based on the GK-EM shifts, the estimated average negative surface charge spacing near the KD channel is 39 A. To quantify the effects of protons on the rates of transitions between the hypothesized conformational states of the channels, we fitted the experimental macroscopic steady state conductance-voltage relationship and the voltage dependence of time constants of activation and deactivation, simultaneously, with a sequential three-state model CCO. In terms of this model, protonation affects the voltage-dependent properties via a decrease in localized, rather than homogeneous, surface charge sensed by the gating moieties. In terms of either the CO or CCO model, the protonation of a site with a pKa of 4.8 decreases the voltage-independent number of channels, N, that are available for activation by depolarization.     

Effects of day-to-day changes in root temperature on leaf conductance to water vapour and CO2 assimilation rates of Vigna unguiculata L. Walp.

Summary Leaf gas exchange of Vigna unguiculata was influenced by short-term (day-to-day) changes in soil temperature and the response depended upon the aerial environment. When aerial conditions were constant at 30° C leaf temperature, high air humidity and moderate quantum flux, CO₂ assimilation rate and leaf conductance increased with increases in soil temperature from 20 to 35° C, and this response was reversible. Decreases in CO₂ assimilation rate and leaf conductance were observed at root temperatures above 30° C when root temperatures were increased from 20° C to 40° C and when air humidity was decreased in steps during the day. In contrast, varying soil temperatures between 20 to 35° C had no influence on gas exchange when shoots were subjected to a wide range of temperatures during each day.The gain ratio A/E remained constant at different air humidities when root temperature was less than or equal to 30° C indicating optimal gas exchange regulation, but changed with humidity at higher root temperatures. Leaf conductance responded independently from leaf water potential which remained relatively constant during individual experiments.The results indicate that plant responses to high root temperatures may have relevance to plant performance in semi-arid environments. They also illustrate the importance of controlling soil temperatures when studying the responses of potted plants in controlled aerial environments.Dedicated to K.F. Springer     

Voltage and temperature dependence of single K+ channels isolated from canine cardiac sarcoplasmic reticulum.

The temperature and voltage dependence of gating and conductance of sarcoplasmic reticulum K+ channels (S-R K+) isolated from adult canine hearts were studied using the reconstituted bilayer technique. Fusion of vesicles from this preparation frequently resulted in the incorporation of a single channel. Only bilayers into which a single S-R K+ channel had fused were studied. The three conductance states of the channel, fully open (O2), substate conductance (O1), and closed (C) were studied as a function of voltage (-50 to +50 mV) and temperature (16 to 37 degrees C). Permeation through the O1 state showed the same temperature dependence as the O2 state corresponding to an enthalpy of permeation of 4.1-4.2 kcal/mol, which is similar to that for K+ diffusion through water. As expected, increased temperature increased the frequency of gating transitions and shortened the average dwell time spent in any conductance state. Over the range of 25 to 37 degrees C, the average dwell time spent in the O1, O2, and C states decreased by 44 +/- 11, 36 +/- 13, and 78 +/- 7% (n = 3 to 4 channels), respectively. The ratio of probabilities between the various conductance states was not strongly temperature sensitive. Analysis of the voltage dependence of this channel was carried out at 37 degrees C and revealed that the dwell times of the O1 and O2 states were voltage insensitive and the probability ratio (PO2:PO1) was approximately 7 and was voltage insensitive.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of High Temperature on Photosynthesis in Beans (I. Oxygen Evolution and Chlorophyll Fluorescence)

We studied the effect of increasing temperature on photosynthesis in two bean (Phaseolus vulgaris L.) varieties known to differ in their resistance to extreme high temperatures, Blue Lake (BL), commercially available in the United Kingdom, and Barbucho (BA), noncommercially bred in Chile. We paid particular attention to the energy-transducing mechanisms and structural responses inferred from fluorescence kinetics. The study was conducted in non-photorespiratory conditions. Increases in temperature resulted in changes in the fluorescence parameters nonphotochemical quenching (qN) and photochemical quenching (qP) in both varieties, but to a different extent. In BL and BA the increase in qP and the decrease in qN were either completed at 30[deg]C or slightly changed following increases from 30 to 35[deg]C. No indication of photoinhibition was detected at any temperature, and the ratio of the quantum efficiencies of photosystem II (PSII) and O2 evolution remained constant from 20 to 35[deg]C. Measurements of 77-K fluorescence showed an increase in the photosystem I (PSI)/PSII ratio with temperature, suggesting an increase in the state transitions. In addition, measurements of fast-induction fluorescence revealed that the proportion of PSII[beta] centers increased with increasing temperatures. The extent of both changes were maximum at 30 to 35[deg]C, coinciding with the ratio of rates at temperatures differing by 10[deg]C for oxygen evolution.     

The Effect of Leaf Temperature and Photorespiratory Conditions on Export of Sugars during Steady-State Photosynthesis in Salvia splendens

Export and photosynthesis in leaves of Salvia splendens were measured concurrently during steady-state 14CO2 labeling conditions. Under ambient CO2 and O2 conditions, photosynthesis and export rates were similar at 15 and 25[deg]C, but both declined as leaf temperature was raised from 25 to 40[deg]C. Suppressing photorespiration between 15 and 40[deg]C by manipulating CO2 and O2 levels resulted in higher rates of leaf photosynthesis, total sugar synthesis, and export. There was a linear relationship between the rate of photosynthesis and the rate of export between 15 and 35[deg]C. At these temperatures, 60 to 80% of the carbon fixed was readily exported with sucrose, raffinose, and stachyose, which together constituted over 90% of phloem mobile assimilates. Above 35[deg]C, however, export during photosynthesis was inhibited both in photorespiratory conditions, which inhibited photosynthesis, and in nonphotorespiratory conditions, which did not inhibit photosynthesis. Sucrose and raffinose but not stachyose accumulated in the leaf at 40[deg]C. When leaves were preincubated at 40[deg]C and then cooled to 35[deg]C, export recovered more slowly than photosynthesis. These data are consistent with the view that impairment of export processes, rather than photosynthetic processes associated with light trapping, carbon reduction, and sucrose synthesis, accounted for the marked reduction in export between 35 and 40[deg]C. Taken together, the data indicated that temperature changes between 15 and 40[deg]C had two effects on photosynthesis and concurrent export. At all temperatures, suppressing photorespiration increased both photosynthesis and export, but above 35[deg]C, export processes were more directly inhibited independent of changes in photorespiration and photosynthesis.     

Effect of High Temperature on Photosynthesis in Beans (II. CO2 Assimilation and Metabolite Contents)

The effect of high temperatures on CO2 assimilation, metabolite content, and capacity for reducing power production in non-photorespiratory conditions has been assessed in two different bean (Phaseolus vulgarus L.) varieties, Blue Lake (commercially available in the United Kingdom) and Barbucho (a noncommercially bred Chilean variety), which are known to differ in their resistance to extreme high temperatures. Barbucho maintains its photosynthetic functions for a longer period of time under extreme heat compared with Blue Lake. The CO2 assimilation rate was increased by increases in temperature, with a decrease in ratio of rates of temperatures differing by 10[deg]C. It is suggested that limitations to CO2 assimilation are caused by metabolic restrictions that can be differentiated between those occurring in the range of 20 to 30[deg]C and 30 to 35[deg]C. It is likely that changes in the capacity for Calvin cycle regeneration and starch synthesis affect photosynthesis in the range of 20 to 30[deg]C. But following an increase in temperature from 30 to 35[deg]C, the supply of reducing power becomes limiting. From analysis of adenylate concentration, transthylakoid energization, and, indirectly, NADPH/NADP+ ratio, it was concluded that the limitation in the assimilatory power was due to an oxidation of the NADPH/NADP+ pool. In the range of 30 to 35[deg]C, the photosystem I quantum yield increased and photosystem II maintained its value. We conclude that the reorganization of thylakoids observed at 30 to 35[deg]C increased the excitation of photosystem I, inducing an increase in cyclic electron transport and a decrease in the supply of NADPH, limiting carbon assimilation.     

Dual regulation of the n type K+ channel in Jurkat T lymphocytes by protein kinases A and C.

We have measured the activity of the n type K+ channel present in human (Jurkat) T lymphocytes using the patch clamp technique in the whole-cell configuration. We report that protein kinase A (PKA) and protein kinase C (PKC) modulate, in a dual manner, the K+ conductance in these cells. Activation of PKA decreases the amplitude of the current, as previously reported (Bastin, B., Payet, M. D., and Dupuis, G. (1990) Cell. Immunol. 128, 385-399), and this is also the case for 12-O-tetradecanoylphorbol-13-acetate-dependent activation of PKC. In contrast, inhibitors of PKC (H7, staurosporine, polymixin B, and anti-PKC antibody) increase the current amplitude. Of importance, down-regulation of PKC or its inhibition prevented the PKA-dependent inhibition of the K+ channels. Addition of alkaline phosphatase via the patch pipette increased the K+ conductance under basal conditions and reversed the inhibition produced by PKA. The dual modulation of K+ channels in Jurkat T cells is in agreement with the presence of consensus sequences in the primary structure of the n type K+ channel.     

Exogenous Abscisic Acid Mimics Cold Acclimation for Cacti Differing in Freezing Tolerance

The responses to low temperature were determined for two species of cacti sensitive to freezing, Ferocactus viridescens and Opuntia ficus-indica, and a cold hardy species, Opuntia fragilis. Fourteen days after shifting the plants from day/night air temperatures of 30/20[deg]C to 10/0[deg]C, the chlorenchyma water content decreased only for O. fragilis. This temperature shift caused the freezing tolerance (measured by vital stain uptake) of chlorenchyma cells to be enhanced only by about 2.0[deg]C for F. viridescens and O. ficus-indica but by 14.6[deg]C for O. fragilis. Also, maintenance of high water content by injection of water into plants at 10/0[deg]C reversed the acclimation. The endogenous abscisic acid (ABA) concentration was below 0.4 pmol g-1 fresh weight at 30/20[deg]C, but after 14 d at 10/0[deg]C it increased to 84 pmol g-1 fresh weight for O. ficus-indica and to 49 pmol g-1 fresh weight for O. fragilis. Four days after plants were sprayed with 7.5 x 10-5 M ABA at 30/20[deg]C, freezing tolerance was enhanced by 0.5[deg]C for F. viridescens, 4.1[deg]C for O. ficus-indica, and 23.4[deg]C for O. fragilis. Moreover, the time course for the change in freezing tolerance over 14 d was similar for plants shifted to low temperatures as for plants treated with exogenous ABA at moderate temperatures. Decreases in plant water content and increases in ABA concentration may be important for low-temperature acclimation by cacti, especially O. fragilis, which is widely distributed in Canada and the United States.     

Outwardly rectifying chloride channels in lymphocytes

Summary Outwardly rectifying Cl^– channels in cultured human Jurkat T-lymphocytes were activated by excising a patch of membrane using the inside-out (i/o) patch-clamp configuration and holding at depolarized voltages for prolonged periods of time (1–6 min at +80 mV, 20°C). The single-channel current at +80 mV was 4.5 ± 0.3 pA and at –80 mV, it was 1.0 ± 0.4 pA. After activation, the probability of being open (P ₀)for the lymphocyte channel was voltage independent. Activation of the Cl^– channel in lymphocytes was temperature dependent. Nineteen percent of i/o recordings from lymphocytes made at 20°C exhibited Cl^– channel activity. In contrast, 49% of recordings made at 30°C showed channel activity. The number of channels in an active patch was not significantly different at the two temperatures. Channel activation in excised, depolarized patches also occurred 20-fold faster at 30°C than at 20°C. There was no marked change in the single-channel conductance at 30°C. Open-channel conductance was blocked by 200 m indanyloxyacetic acid (IAA) or 1 mm SITS when applied to the intracellular side of the patch. The characteristics of this channel are similar to epithelial outwardly rectifying Cl^– channels thought to be involved in fluid secretion     

Differential effects of temperature on E. coli and synthetic polyhydroxybutyrate/polyphosphate channels

Complexes of poly-(R)-3-hydroxybutyrate and inorganic polyphosphate (PHB/polyP), isolated from the plasma membranes of Escherichia coli or prepared synthetically (HB(128)/polyP(65)), form Ca(2+)-selective ion channels in planar lipid bilayers that exhibit indistinguishable gating and conductance characteristics at 22 degrees C. Here we examine the gating and conductance of E. coli and synthetic PHB/polyP complexes in planar lipid bilayers as a function of temperature from 15 to 45 degrees C. E. coli PHB/polyP channels remained effectively open throughout this range, with brief closures that became more rare at higher temperatures. Conversely, as temperatures were gradually increased, the open probability of HB(128)/polyP(65) channels progressively decreased. The effect was fully reversible. Channel conductance exhibited three distinct phases. Below 25 degrees C, as PHB approached its glass temperature (ca. 10 degrees C), the conductance of both E. coli and synthetic channels remained at about the same level (95-105 pS). Between 25 degrees C and ca. 40 degrees C, the conductance of E. coli and synthetic channels increased gradually with temperature coefficients (Q(10)) of 1.45 and 1.42, respectively. Above 40 degrees C, E. coli channel conductance increased sharply, whereas the conductance of HB(128)/polyP(65) channels leveled off. The discontinuities in the temperature curves for E. coli channels coincide with discontinuities in thermotropic fluorescence spectra and specific growth rates of E. coli cells. It is postulated that E. coli PHB/polyP complexes are associated with membrane components that inhibit their closure at elevated temperatures.