Donation of Electrons from Cytosolic Components to the Intersystem Chain in the Cyanobacterium Synechococcus sp. PCC 7002 as Determined by the Reduction of P700+

Cells, of Synechococcus sp. PCC 7002 showed a low oxidationlevel of P700 under a far-red light at 6 W m^–2 which inducednearly complete oxidation of P700 in spinach leaves, and a strongerfar-red light was required to observe the oxidation of P700.DCMU did not affect the level of P700⁺² but 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinoneinduced the oxidation of P700 under far-red light, indicatingthat the low oxidation level of P700 was due to the donationof electrons to P700⁺² from the cytosolic respiratory donorsthrough the intersystem chain at the plastoquinone pool. Theelectron transfer from the cytosolic donors to the intersystemchain was inhibited by HgCl₂ but not by antimycin A. The reductionof P700⁺ in Synechococcus cells, after illumination by strongfar-red light was mostly accounted for by the electron flowto the inter system chain from the respiratory donors (t     

Reaction of Reconstituted Acceptor Quinone and Dynamic Equilibration of Electron Transfer in the Photosystem I Reaction Center

Electron transfer mechanism in the spinach photosystem I reactioncenter that contains artificial quinones in place of phylloquinone(2-methyl-3-phytyl-1,4-naphthoquinone, vitamin K₁) as the secondaryelectron acceptor, Q_ø (or A₁) was discussed. (1) Mostof the reconstituted quinones oxidized the primary acceptorchlorophyll a, A⁰, at a rate rapid enough to compete againstthe charge recombination between A₀^– and the oxidizeddonor chlorophyll P700⁺. (2) The pathway of electron transferfrom the semiquinone^– varied depending on the redox potentialvalue of each semiquinone^– /quinone couple. Low potentialquinones reduced the tertiary acceptor iron-sulfur center, F_x,while the high potential ones reduced P700⁺ directly with a200-µs halftime. (3) The E_m value of each semiquinone^–/quinone couple in situ in the reaction center was estimatedto be shifted by about 0.3 volt to the negative side from theirhalf wave redox potential values that were measured polarographicallyin dimethylformamide. The shift seems to represent the acceptorproperty of the protein environment at the Q_ø site. (4)The E_m of reconstituted phylloquinone was estimated to be 50–80mV more negative than that of F_x. (5) The mechanism of efficientelectron transfer in the reaction center was discussed basedon the dynamic equilibria between the electron transfer componentsand on the estimated E_m values. (Received April 9, 1994; Accepted July 7, 1994)     

Targeted inactivation of the gene psaK encoding a subunit of photosystem I from the cyanobacterium Synechocystis sp. PCC 6803

Mutant strains of the unicellular cyanobacterium Synechocystissp. PCC 6803, in which the psaK gene was insertionally inactivatedby targeted mutagenesis, were constructed. The gene is one ofthe two potential PsaK-coding genes which have been found asa result of the genome project with this cyanobacterium. Oneof the mutants was characterized in detail. A monocistronic,480-nucleotide mRNA of psaK was absent in total RNA from themutant cells. Inactivation of psaK had little effect on theaccumulation of polypeptides in the isolated PSI complexes exceptfor a polypeptide with an apparent molecular mass of 4.6 kDawhich was absent in the mutant. The amino-terminal amino acidsequence of the 4.6-kDa polypeptide confirmed that it was thetranslation product of psaK and further revealed a presequenceof PsaK. Characteristics of photoautotrophic growth at differenttemperatures, the amount of chlorophyll per cell, photosyntheticelectron transport rates with various electron acceptors, thekinetics of charge recombination between P700⁺ and reduced F_A/F_B,and the molar ratio of chlorophyll to P700, of the mutant werenot significantly different from those of the wild type. Furthermore,the trimer to monomer ratio of the PSI complexes isolated fromthe mutant was similar to that isolated from the wild type. (Received July 27, 1998; Accepted October 13, 1998)     

Dibromothymoquinone (DBMIB) Replaces the Function of QA at 77 K in the Isolated Photosystem II Reaction Center (D1-D2-Cytochrome b559) Complex: Difference Spectrum of the P680+(DBMIB-) State

Transient absorbance changes of the primary electron donor chlorophylla (P680) and acceptor pheophytin a (H) were measured at 77 Kby nanosecond laser spectroscopy in the D1-D2-cytochrome b₅₅₉photosystem II reaction center complex containing dibromomethylisopropylbenzoquinone (DBMIB). After the laser excitation of the reactioncenter in the presence of DBMIB, only the P680⁺-(DBMIB^-) statewas detected. P680⁺ mainly decayed with a t_1/e of 11 ms. Inthe absence of DBMIB, the excitation produced the P680⁺H^- radicalpair. The radical pair produced the triplet state (P680^T) witha t_1/e of 50 ns, and P680^T then decayed with a t_1/e of 2.1 ms.It was concluded that H_- was oxidized by DBMIB in a time rangefaster than the detecting time resolution (3.5 ns) even at 77K. The rapid oxidation of H^- by DBMIB was also confirmed bythe suppression of delayed fluorescence with a decay t_1/e of50 ns. The P680⁺(DBMIB^-)/P680(DBMIB) difference spectrum exhibiteda Q_y, band with a peak at 682 nm with a shoulder at 673 nm.The spectral shape was almost temperature insensitive between77 and 265 K. The feature of this spectrum in the wavelengthrange between 330 and 720 nm was compared with that of P680_T/P680or H^-/H at 77 K. (Received May 8, 1996; Accepted June 24, 1996)     

Intracellular Cs+ activates the PKA pathway,revealing a fast,reversible, Ca2+-dependent inactivation of L-type Ca2+ current

Inactivation of the L-type Ca²⁺ current (I_CaL) was studied in isolated guinea pig ventricular myocytes with different ionic solutions. Under basal conditions, I_CaL of 82% of cells infused with Cs⁺-based intracellular solutions showed enhanced amplitude with multiphasic decay and diastolic depolarization-induced facilitation. The characteristics of I_CaL in this population of cells were not due to contamination by other currents or an artifact. These phenomena were reduced by ryanodine, caffeine, cyclopiazonic acid, the protein kinase A inhibitor H-89, and the cAMP-dependent protein kinase inhibitor. Forskolin and isoproterenol increased I_CaL by only 60% in these cells. Cells infused with either N-methyl-D-glucamine or K⁺-based intracellular solutions did not show multiphasic decay or facilitation under basal conditions. Isoproterenol increased I_CaL by 200% in these cells. In conclusion, we show that multiphasic inactivation of I_CaL is due to Ca²⁺-dependent inactivation that is reversible on a time scale of tens of milliseconds. Cs⁺ seems to activate the cAMP-dependent protein kinase pathway when used as a substitute for K⁺ in the pipette solution. L-type calcium current; calcium-dependent inactivation; facilitation; phosphorylation; cesium     

Ion Effects on the H+-Translocating Adenosine Triphosphatase and Pyrophosphatase Associated with the Tonoplast of Chora corallina

H⁺-translocating ATPase and pyrophosphatase (PPase) associatedwith the tonoplast of Chara corallina were isolated with theaid of a perfusion technique, and the effects of ions on theiractivities were studied. All the alkali metal cations testedstimulated the ATPase and ATPdependent H⁺ pumping activitiesonly by 10 to 40%. Anions, on the other hand, strongly affectedthe activities. Potassium salts of Cl^- and Br^- stimulated them,while F^- and NO₃^- inhibited them. By contrast, the H⁺-translocatingPPase was insensitive to anions but sensitive to cations. Theorder of cation stimulation was Rb⁺=K⁺>Cs⁺>Na⁺=Li⁺>choline⁺.NO₃^- (50 mil), thought to be a specific inhibitor of the tonoplast-typeH⁺-ATPase, inhibited the ATPdependent H⁺ pumping almost completelybut the ATPase activity by only about 50%. Na⁺ inhibited thePP₁-dependent H⁺ pumping (I_5O=5OmM) in the presence of 50 mMKCl but not the ATP-dependent one. The PPase was more sensitiveto F^- (I₅₀=400µM) than the ATPase. Both the H⁺-ATPaseand the H⁺-PPase required Mg²⁺ for their activities, althoughan excess was inhibitory to both. The different sensitivitiesof the PP₁-dependent and the ATP-dependent H⁺- pumping enzymesto ions correspond to the tonoplast enzymes of higher plantsand may be used as "markers" to distinguish between these enzymesin characean cells (Received October 2, 1987; Accepted May 18, 1988)     

Cl- currents activated via purinergic receptors in Xenopus follicles

Ionic currents elicited via purinergic receptors located in themembrane of Xenopus follicles werestudied using electrophysiological techniques. Follicles responded toATP-activating inward currents with a fast time course(F_in). InRinger solution, reversal potential (E_rev) ofF_in was 22mV, which did not change with external substitutions ofNa⁺ orK⁺, whereas solutions containing50 or 5% of normal Clconcentration shiftedE_rev to about +4and +60 mV, respectively, and decreasedF_in amplitude,indicating thatF_in was carriedby Cl.F_in had an onsetdelay of ~400 ms, measured by application of a brief jet of ATP froma micropipette positioned near the follicle (50 µm).F_in was inhibitedby 50% in follicles pretreated with pertussis toxin. This suggests a Gprotein-mediated receptor channel pathway.F_in was mimickedby 2-MeSATP and UTP, the potency order (half-maximal effectiveconcentration) was 2-MeSATP (194 nM) > UTP (454 nM) > ATP(1,086 nM). All agonists generatedCl currents and displayedcross-inhibition on the others.F_in activation byacetylcholine also cross-inhibitedF_in-ATPresponses, suggesting that all act on a common channel-activationpathway.

     

影响叶螨磷酸酯酶活性的四因子数学模型

应用二次回归通用旋转组合设计,组建了影响叶螨磷酸酯酶（酸性和碱性）活性的四因子（缓冲液Ph值X₁、温浴时间X₂、反应温度X₃、底物浓度X₄）数学模型: Y_酸性=0.456380+0.107889X₂+0.069027X₃-0.026836X₁²-0.030794X₃², F=24.98,P<0.01;Y_碱性=0.267286-0.200736X₁+0.049541X₂+0.030930X₃-.049063X₁X₂+0.053585X₁²-0.049665X₂^2, F=57.68,P<0.01。结果表明,温浴时间是影响叶螨酸性磷酸酯酶活性的关键因子,在缓冲液pH 4.4、底物浓度8.5×10^-3 mol/L、42℃温浴40 min测得该酶活性最强。影响碱性磷酸酯酶活性的关键因子则是缓冲液pH值,pH 9.0、37℃恒温30 min、底物7.5×10^-3 mol/L的条件下,光密度值最大。两种酶的最大吸收峰波长为405 nm。     

Amino Acid Sequence Similarities between the Vacuolar Proton-Pumping Inorganic Pyrophosphatase and the c-Subunit of F0F1-ATPases

Comparison of the Arabidopsis thaliana vacuolar proton-pumpinginorganic pyrophosphatase with three F₀F₁-ATPase c-subunitsrevealed a strong similarity between a stretch containing aminoacids 227–245 of the H⁺-PPase and a transmembrane a-helixof the c-subunits which contains the glutamate which binds N,N'-dicyclohexylcarbodiimide. (Received November 16, 1992; Accepted December 22, 1992)     

Effects of Seed Treatments on the Emergence of Oil Seed Rape Seedlings from Compacted Soil

Imbibition of seeds of oil seed rape (Brassica napus cv Jetneuf)in 10^–3 M aminoethoxyvinylglycine (AVG) or 10^–2silver thiosulphate (Ag⁺) had no effect on germination nor onthe emergence of seedlings from uncompacted or lightly compressedsoil, but significantly reduced emergence from moderately compressedsoil of 68.4 or 143.3 N cm^–2 impedance. Exertion of force by emerging control seedlings against a staticcantilever bar fitted with strain gauges reached a maximum (F_max)of 6 g over 10 h. Higher axial forces were achieved when theseedlings were emerging from compressed soil, without any changein the time required to reach F_max, so that the build-up offorce was considerably (1.8 fold) faster than in uncompressedsoil. This adaptive response to soil impedance was modified by theseed pretreatments employed. Seedlings from AVG or Ag⁺pretreatedseeds produced lower axial forces than controls, and neitherF_max nor the rate at which force developed showed any responseto soil compression. After pretreatment in 10^–3 ethephon or 10^–4 naphthaleneacetic acid (NAA) the seedlings achieved similar F_max to controlseedlings, but responded more rapidly to soil compression sothat the rate of build up of emergence force was 2.3 fold (NAA)or 2.8 fold (ethephon) faster in compressed than in uncompressedsoil. The results suggest that the exertion of force by a seedlingagainst a barrier involves a dynamic response to impedance onthe part of the seedling. This response can be either enhancedor suppressed by substances which affect ethylene productionor ethylene action. Such compounds may have promise for modifyingseedling emergence from impeding soils. Brassica napus, oil seed rape, seedling emergence, soil compaction, ethylene, Ethrel, silver, aminoethoxyvinylglycine, naphthalene acetic acid     

Isolation of the Photoactive Reaction Center Complex that Contains Three Types of Fe-S Centers and a Cytochrome c Subunit from the Green Sulfur Bacterium Chlorobium limicola f. thiosulfatophilum, Strain Larsen

The photoactive reaction center (RC) complex from the greensulfur bacterium Chlorobium limicola f. thiosulfatophilum, strainLarsen, was isolated after solubilization and ammonium sulfatefractionation followed by ion-exchange chromatography. The spectrumof the complex was almost identical with that of the similarRC complex isolated by Feiler et al. [(1992) Biochemistry 31:2608–2614] except for the presence of cytochrome c₅₅₁instead of c₅₅₃ in the latter study. A molecular ratio of BChla to P840 of the isolated RC complex was assayed to be 25–35.SDSPAGE analysis revealed that the isolated complex containedthree major polypeptides with apparent molecular masses of 68,41 and 21 kDa, respectively. The 21-kDa polypeptide was identifiedto be a heme-binding protein by staining the gel for peroxidaseactivity. The cytochrome c₅₅₁ was oxidized by flash light ina biphasic manner with half times of 90 and 390 µs, respectively,that coincided with the reduction half times of P840⁺. Threedistinct iron-sulfur centers assigned to F_A, F_B and F_x, respectively,from their g-values were detected by EPR spectroscopy at cryogenictemperature. These results suggest that the present preparationcontains a minimal functional unit of the RC of this bacterium,and that this complex appears to lie on a evolutionary linebetween RC's of purple bacteria and photosystem I. (Received August 18, 1992; Accepted October 28, 1992)     

The Steady State Chlorophyll a Fluorescence Exhibits in Vivo an Optimum as a Function of Light Intensity which Reflects the Physiological State of the Plant

Modulated (690 and 730 nm), as well as direct chlorophyll (Chl)a fluorescence and changes in the concentration of the oxidizedP700 were measured under steady state conditions in leaves ofhigher plants adapted to different light intensities. All theleaf samples exhibit an optimum curve of steady state fluorescenceyield (F_s) versus the light intensity but its position withrespect to light intensity varies considerably from one speciesto another or from one sample to other even in the same plantor within the same leaf sample. However, the optimum level ofF_s was always at a moderate light intensity. By using the modulatedfluorescence technique, the system with all closed (F^l_m) oropen reaction center (F^l_o) were measured in steady state conditions.Each experimentally measured fluorescence yield was separatedinto a fluorescence emission of open (F_open = F^l_o,(1—V_s))and closed (F_closed = (F^l_m . V_s)) reaction center (RC) of photosystemII where V_s=(F_s – F^l_o)/(F^l_m – F^l_o) is the functionof fraction of closed reaction centers. With increasing lightintensity, the fraction of open RC decreased while the fractionof closed RC increased. Maximum quantum efficiency (P_o) andactual quantum efficiency (P) decreased by increasing lightintensity. An optimum level of F_s was observed, when the fractionof closed reaction centers V_s of each sample was about 0.2 showinga common quenching mechanism which determines the fluorescenceproperties under steady state condition. This explains the apparentphenomenological contradiction that the fluorescence yield understeady state conditions can increase or decrease upon an increaseof actinic light. (Received December 31, 1994; Accepted May 1, 1995)     

Time-Resolved Spectroscopy of Chlorophyll-a like Electron Acceptor in the Reaction Center Complex of the Green Sulfur Bacterium Chlorobium tepidum

The absorption changes of chlorophyll (Chl) a-like pigments(C670) were studied by ns-ms laser spectroscopy at 77 K in theuntreated and urea-treated homodimeric reaction center (RC)complex of the green sulfur bacterium Chlorobium tepidum. Theuntreated RC complex contained 9 molecules of C670 in additionto 41 molecules of Bchl a and 0.9 molecules of menaquinone-7per one primary electron donor Bchl a dimer (P840). Upon photo-oxidationof P840, C670 showed an absorption change of a red-shift withan isosbestic wavelength at 668 nm. The absorption change ofP840 decayed with time constants (t_1/e) of 55 and 37 ms at 283and 77 K, respectively, and was assigned to represent the chargerecombination between P840⁺ and FeS^–. In the urea-treatedRC complex, a bleach peaking at 670 nm with a shoulder peakat 662 nm, which is ascribable to the reduced primary electronacceptor A₀^–, was detected after the laser excitationin addition to the shift at 668 nm indicating the formationof the P840⁺A₀^– state. The P840⁺A₀^– state decayedwith a t_1/e of 43 ns at 77 K and produced a triplet state p840^Tdue to the suppression of the forward electron transfer. Theseresults indicate the two different types of C670 species inthe RC complex; the one peaking at 670 nm functions as A⁰, whilethe other peaking at 668 nm shows the electrochromic shift,which presumably functions as the accessory pigment locatedin the close vicinity of P840. (Received May 17, 1999; Accepted July 14, 1999)     

Pool Size of Electrons That Can Be Donated to P700+ As Determined in Intact Leaves: Donation to P700+ from Stromal Components Via the Intersystem Chain

A method for estimation of the functional pool size of electronsthat can be donated to P700⁺ after the illumination of intactleaves with actinic light is described. The complementary areasbetween the stationary level of P700⁺ attained by irradiationwith far-red light and the oxidation curves of P700 by far-redlight after a 50-ms multiple-turnover light (MT-area), and afterthe illumination of actinic light (AL-area) were determined.Since the MT-area represents the pool size of electrons in theintersystem chain, the ratio of the AL-area to the MT-area allowsus to estimate the pooi size of electrons stored during actinicillumination that can be donated to P700⁺ The ratio of the AL-areato the MT-area was determined for intact leaves of several C₃plants, and it was found to increase to a value of two or threewith increases in the intensity of actinic light, and it wasfurther increased by anaerobiosis. The pool size of electronsthat can be donated to P700⁺ from the stroma was estimated tobe in the range of 12 and 28 electrons per P700 under aerobicconditions. These results indicate that stromal components donateelectrons to P700⁺ through the intersystem chain in chloroplastsof C₃ plants. (Received July 3, 1992; Accepted September 29, 1992)     

Mechanism of shortened action potential duration in Na+-Ca2+ exchanger knockout mice

In cardiac-specific Na⁺-Ca²⁺ exchanger (NCX) knockout (KO) mice, the ventricular action potential (AP) is shortened. The shortening of the AP, as well as a decrease of the L-type Ca²⁺ current (I_Ca), provides a critical mechanism for the maintenance of Ca²⁺ homeostasis and contractility in the absence of NCX (Pott C, Philipson KD, Goldhaber JI. Excitation-contraction coupling in Na⁺-Ca²⁺ exchanger knockout mice: reduced transsarcolemmal Ca²⁺ flux. Circ Res 97: 1288–1295, 2005). To investigate the mechanism that underlies the accelerated AP repolarization, we recorded the transient outward current (I_to) in patch-clamped myocytes isolated from wild-type (WT) and NCX KO mice. Peak I_to was increased by 78% and decay kinetics were slowed in KO vs. WT. Consistent with increased I_to, ECGs from KO mice exhibited shortened QT intervals. Expression of the I_to-generating K⁺ channel subunit Kv4.2 and the K⁺ channel interacting protein was increased in KO. We used a computer model of the murine AP (Bondarenko VE, Szigeti GP, Bett GC, Kim SJ, and Rasmusson RL. Computer model of action potential of mouse ventricular myocytes. Am J Physiol Heart Circ Physiol 287: 1378–1403, 2004) to determine the relative contributions of increased I_to, reduced I_Ca, and reduced NCX current (I_NCX) on the shape and kinetics of the AP. Reduction of I_Ca and elimination of I_NCX had relatively small effects on the duration of the AP in the computer model. In contrast, AP repolarization was substantially accelerated when I_to was increased in the computer model. Thus, the increase in I_to, and not the reduction of I_Ca or I_NCX, is likely to be the major mechanism of AP shortening in KO myocytes. The upregulation of I_to may comprise an important regulatory mechanism to limit Ca²⁺ influx via a reduction of AP duration, thus preventing Ca²⁺ overload in situations of reduced myocyte Ca²⁺ extrusion capacity. genetically altered mice; cardiac myocytes; short QT interval; transient outward current     

Dependence of Na+-K+ pump current-voltage relationship on intracellular Na+, K+, and Cs+ in rabbit cardiac myocytes

To examine effects of cytosolicNa⁺, K⁺, and Cs⁺ on the voltagedependence of the Na⁺-K⁺ pump, we measuredNa⁺-K⁺ pump current (I_p)of ventricular myocytes voltage-clamped at potentials(V_m) from 100 to +60 mV. Superfusates weredesigned to eliminate voltage dependence at extracellular pump sites.The cytosolic compartment of myocytes was perfused with patch pipette solutions with a Na⁺ concentration ([Na]_pip)of 80 mM and a K⁺ concentration from 0 to 80 mM or withsolutions containing Na⁺ in concentrations from 0.1 to 100 mM and K⁺ in a concentration of either 0 or 80 mM. When[Na]_pip was 80 mM, K⁺ in pipette solutionshad a voltage-dependent inhibitory effect on I_pand induced a negative slope of theI_p-V_m relationship. Cs⁺ in pipette solutions had an effect onI_p qualitatively similar to that ofK⁺. Increases in I_p with increasesin [Na]_pip were voltage dependent. The dielectriccoefficient derived from[Na]_pip-I_p relationships at thedifferent test potentials was 0.15 when pipette solutions included 80 mM K⁺ and 0.06 when pipette solutions were K⁺ free.

     

Changes of action potentials and force at lowered [Na+]o in mouse skeletal muscle: implications for fatigue

We examined 1) whether the effects of lowered trans-sarcolemmal Na⁺ gradient on force differed between nonfatigued fast- and slow-twitch muscles of mice and 2) whether effects on action potentials could explain the decrease of force. The Na⁺ gradient was reduced by lowering the extracellular [Na⁺] ([Na⁺]_o). The peak force-[Na⁺]_o relationships for the twitch and tetanus were the same in nonfatigued extensor digitorum longus and soleus muscles: force was maintained over a large range of [Na⁺]_o and then decreased abruptly over a much smaller range. However, fatigue was significantly exacerbated at a lowered [Na⁺]_o that had little effect in nonfatigued soleus muscle. This finding suggests that substantial differences exist in the Na⁺ effect on force between nonfatigued and fatigued muscle. The reduced contractility in nonfatigued muscles at lowered [Na⁺]_o was largely due to 1) an increased number of inexcitable fibers and threshold for action potentials, 2) a reduction of action potential amplitude, and 3) a reduced capacity to generate action potentials throughout trains. sodium gradient; muscle contraction; action potential train; extensor digitorum longus; soleus     

Picosecond Photochemistry of P700-Enriched and Vitamin K1-Depleted Photosystem I Particles Isolated from Spinach

Picosecond transient absorption changes, with a laser intensityas low as one photon absorbed per single reaction center, weremeasured with vitamin K₁-depleted and P700-enriched particleswhich were obtained by ether treatment of spinach PS-I particles.When P700 was in the oxidized state, a bleaching that correspondedto about one-seventh of the ground state absorption was observedjust after a laser flash (0 picosecond delay). A major partof the bleaching decayed with a lifetime of about 35 picoseconds,which corresponds to the relaxation of the excited antenna chl-ato the ground state. By contrast, when P700 was in the reducedstate, the bleaching observed at a 0 ps delay was broader, especiallyon the longer wavelength side than the ground state absorption,probably because of the generation of the excited state of P700.About one half of the bleaching decayed within 35 ps and theremaining half, which had a broad spectrum and a peak around682 nm, was conserved up to 2 ns. This long-lived bleachingprobes no picosecond decay of the radical pair P700⁺-A₀^–because electrons were not transferred from A₀¹ to A₁ in vitaminK₁-depleted particles. After addition of vitamin K₃, an analogof vitamin K₁, to the reduced particles, the bleaching around685 nm decayed successively with an apparent rate of about 150picosecond, while the bleaching around 700 nm was conservedfor up to 2 nanosecond. Thus, the bleaching remaining at 2 nsresembled the difference spectrum of P700, suggesting a subnanosecondquenching of A₀¹ by the externally added vitamin K₃. These observationssupport a recent proposal that the secondary electron acceptorA₁, in photosystem I, is vitamin K₁. ³Permanent address: Optics Laboratory, Korea Standards ResearchInstitute, Daedok Science Town, Chungnam 300-31, Korea. (Received October 24, 1988; Accepted April 14, 1989)     

Differences in nitrate and ammonia uptake among components of a phytoplankton population

We examined the NO₃^– and NH₄⁺ uptake capabilities of thedinoflagellate Peridinium cinctum and of the accompanying nanoplanktonduring the spring P. cinctum bloom in Lake Kin-neret. Throughoutthe Peridinium season, the smaller algae had greater affinitiesand faster specific uptake rates for both NO₃^–and NH₄⁺It appears that P. cinctum cannot directly compete with nanoplanktonfor nitrogen nutrients. Other factors such as the ability ofdinoflagellates to swim freely in the water column and low grazingpressures may explain their dominance in the lake.     

Involvement of anion channel(s) in the modulation of the transient outward K(+) channel in rat ventricular myocytes

The cardiac Ca²⁺-independent transient outward K⁺ current (I_to), a major repolarizing ionic current, is markedly affected by Cl^– substitution and anion channel blockers. We reexplored the mechanism of the action of anions on I_to by using whole cell patch-clamp in single isolated rat cardiac ventricular myocytes. The transient outward current was sensitive to blockade by 4-aminopyridine (4-AP) and was abolished by Cs⁺ substitution for intracellular K⁺. Replacement of most of the extracellular Cl^– with less permeant anions, aspartate (Asp^–) and glutamate (Glu^–), markedly suppressed the current. Removal of external Na⁺ or stabilization of F-actin with phalloidin did not significantly affect the inhibitory action of less permeant anions on I_to. In contrast, the permeant Cl^– substitute Br^– did not markedly affect the current, whereas F^– substitution for Cl^– induced a slight inhibition. The I_to elicited during Br^– substitution for Cl^– was also sensitive to blockade by 4-AP. The ability of Cl^– substitutes to induce rightward shifts of the steady-state inactivation curve of I_to was in the following sequence: NO₃^– > Cl^– Br^– > gluconate^– > Glu^– > Asp^–. Depolymerization of actin filaments with cytochalasin D (CytD) induced an effect on the steady-state inactivation of I_to similar to that of less permeant anions. Fluorescent phalloidin staining experiments revealed that CytD-pretreatment significantly decreased the intensity of FITC-phalloidin staining of F-actin, whereas Asp^– substitution for Cl^– was without significant effect on the intensity. These results suggest that the I_to channel is modulated by anion channel(s), in which the actin cytoskeleton may be implicated. transient outward potassium current; anion channel; actin cytoskeleton; myocyte; potassium ion