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1.
Kupriyanova E. V. Markelova A. G. Lebedeva N. V. Gerasimenko L. M. Zavarzin G. A. Pronina N. A. 《Microbiology》2004,73(3):255-259
The activity of carbonic anhydrase (CA) was studied in different cell fractions of the alkaliphilic cyanobacterium Microcoleus chthonoplastes. The activity of this enzyme was found in the soluble and membrane protein fractions, as well as in intact cells and in a thick glycocalyx layer enclosing the cyanobacterium cells. The localization of CA in glycocalyx of M. chthonoplastes was shown by western blot analysis and by immunoelectron microscopy studies with antibodies to the thylakoid CA from Chlamydomonas reinhardtii (Cah3). At least one of the CA forms occurring in M. chthonoplastes CA was shown to be an -type enzyme. A possible mechanism of the involvement of the glycocalyx CA in calcification of cyanobacteria is discussed. 相似文献
2.
José de JesÚs Garcia M. Tuena de Gómez-Puyou A. Gómez-Puyou 《Journal of bioenergetics and biomembranes》1995,27(1):127-136
The effect of trifluoperazine (TFP) on the ATPase activity of soluble and paniculate F1ATPase and on ATP synthesis driven by succinate oxidation in submitochondrial particles from bovine heart was studied at pH 7.4 and 8.8. At the two pH. TFP inhibited ATP hydrolysis. Inorganic phosphate protected against the inhibiting action of TFP. The results on the effect of various concentrations of phosphate in the reversal of the action of TFP on hydrolysis at pH 7.4 and 8.8 showed that H2PO
4
–
is the species that competes with TFP. The effect of TFP on oxidative phosphorylation was studied at concentrations that do not produce uncoupling or affect the aerobic oxidation of succinate (<15M). TFP inhibited oxidative phosphorylation to a higher extent at pH 8.8 than at pH 7.4; this was through a diminution in theV
max, and an increase in theK
m for phosphate. Data on phosphate uptake during oxidative phosphorylation at several pH showed that H2PO
4
–
is the true substrate for oxidative phosphorylation. Thus, in both synthesis and hydrolysis of ATP, TFP and H2PO
4
–
interact with a common site. However, there is a difference in the sensitivity to TFP of ATP synthesis and hydrolysis; this is more noticeable at pH 8.8, i.e. ATPase activity of soluble F1 remains at about 40% of the activity of the control in a concentration range of TFP of 40–100M, whereas in oxidative phosphorylation 14M TFP produces a 60% inhibition of phosphate uptake. 相似文献
3.
Summary [14C]Phthalate is transported into L1210 cells via two separate routes, an anion exchange system whose primary substrates are folate compounds, and a second less active system which is sensitive to bromosulfophthalein. When the principal uptake component was blocked by a specific irreversible inhibitor of this system, the remaining route (at pH 7.4) appeared to be saturable and was inhibited by several anions in addition to bromosulfophthalein (K
i
=2 m), including 8-anilino-1-naphthalein sulfonate (K
i
=25 m), unlabeled phthalate (K
i
=500 m), and chloride (K
i
=3500 m). A pronounced effect by pH was also observed. Influx and total uptake of phthalate both increased progressively with decreasing pH and reached values that were 20-fold higher at pH 6.0, compared with pH 7.4. This pH-dependent increase could be blocked, however, by the addition of compounds (nigericin and carbonylcyanidem-chlorophenylhydrazone) which, in combination, collapse proton gradients. Phthalate efflux was relatively insensitive to changes in extracellular pH but could be inhibited (up to 90%) by bromosulfophthalein. Several other anions also inhibited efflux, but to a lesser extent, while chloride, phthalate, lactate, glycolate and acetate enhanced efflux up to 1.8-fold. Efflux also increased at pH 6.0, but not at pH 7.5, upon addition of nigericin and carbonylcyanidem-chlorophenylhydrazone. These results suggest that phthalate is a nonphysiological substrate for a carrier system which mediates transport via an anion/H+ symport mechanism. This system is not the lactate/H+ symport carrier of L1210 cells since: (A) phthalate and lactate influx were inhibited to differing degrees by various anions; and (B) lactic anhydride inhibited the influx and efflux of lactate but had no effect on the transmembrane movement of phthalate. The specificity of this system suggests that its primary anion substrate may be chloride. 相似文献
4.
Z. B. Namsaraev T. V. Kolganova E. O. Patutina D. D. Tsyrenova O. S. Samylina 《Microbiology》2018,87(4):583-590
Clone libraries and morphological analysis were used to investigate cyanobacterial diversity in the cyanobacterial mat and dry crust at the bottom of the shallow, saline, alkaline Lake Khilganta (Southern Siberia, Russia). Filamentous cyanobacteria belonging to Phormidium genus and Coleofasciculus chthonoplastes were found to predominate during the dry period (2006) and the wet periods (1995 and 2012), respectively. Community composition during the dry and wet periods differed significantly. While 11 operational taxonomic units of cyanobacteria were revealed, only 3 occurred during both dry and wet periods. Occurrence of cosmopolitan C. chthonoplastes, which is common in neutral saline environments, is not typical of a continental alkaline lake and may be explained by the similarity of the dominant ions composition in lake water and in seawater. 相似文献
5.
Amaya Blanco-Rivero Tatiana Shutova María José Román Arsenio Villarejo Flor Martinez 《PloS one》2012,7(11)
Background
Cah3 is the only carbonic anhydrase (CA) isoform located in the thylakoid lumen of Chlamydomonas reinhardtii. Previous studies demonstrated its association with the donor side of the photosystem II (PSII) where it is required for the optimal function of the water oxidizing complex. However this enzyme has also been frequently proposed to perform a critical function in inorganic carbon acquisition and CO2 fixation and all mutants lacking Cah3 exhibit very poor growth after transfer to low CO2 conditions.Results/Conclusions
In the present work we demonstrate that after transfer to low CO2, Cah3 is phosphorylated and that phosphorylation is correlated to changes in its localization and its increase in activity. When C. reinhardtii wild-type cells were acclimated to limiting CO2 conditions, the Cah3 activity increased about 5–6 fold. Under these conditions, there were no detectable changes in the level of the Cah3 polypeptide. The increase in activity was specifically inhibited in the presence of Staurosporine, a protein kinase inhibitor, suggesting that the Cah3 protein was post-translationally regulated via phosphorylation. Immunoprecipitation and in vitro dephosphorylation experiments confirm this hypothesis. In vivo phosphorylation analysis of thylakoid polypeptides indicates that there was a 3-fold increase in the phosphorylation signal of the Cah3 polypeptide within the first two hours after transfer to low CO2 conditions. The increase in the phosphorylation signal was correlated with changes in the intracellular localization of the Cah3 protein. Under high CO2 conditions, the Cah3 protein was only associated with the donor side of PSII in the stroma thylakoids. In contrast, in cells grown at limiting CO2 the protein was partly concentrated in the thylakoids crossing the pyrenoid, which did not contain PSII and were surrounded by Rubisco molecules.Significance
This is the first report of a CA being post-translationally regulated and describing phosphorylation events in the thylakoid lumen. 相似文献6.
Erythrocyte membrane potential can be estimated by measuring the transmembrane concentration (activity) distribution of a membrane-permeable ion. We present here the study of difluorophosphate (DFP) as a 19F NMR probe of membrane potential. This bicarbonate and phosphate analogue has a pKa of 3.7±0.2 (SD, n = 4) and therefore exists almost entirely as a monovalent anion at physiological pH. When it is incorporated into red cell suspensions, it gives two well resolved resonances that arise from the intra- and extracellular populations; the intracellular resonance is shifted 130 Hz to higher frequency from that of the extracellular resonance. Hence the transmembrane distribution of DFP is readily assessed from a single 19F NMR spectrum and the membrane potential can be calculated using the Nernst equation. The membrane potential was independent of, DFP concentration in the range 4 to 59 mM, and haematocrit of the cell suspensions of 31.0 to 61.4%. The membrane potential determined by using DFP was 0.94±0.26 of that estimated from the transmembrane pH difference. The distribution ratios of intracellular/extracellular DFP were similar to those of the membrane potential probes, hypophosphite and trifluoroacetate. DFP was found to be transported across the membranes predominantly via the electrically-silent pathway mediated by capnophorin. Using magnetization transfer techniques, the membrane influx permeability-coefficient of cells suspended in physiological medium was determined to be 7.2±2.5 × 10–6 cm s–1 (SD, n=4).
Offprint requests to: P. W Kuchel 相似文献
7.
Thomas Hiltonen Jan Karlsson Kristin Palmqvist Adrian K. Clarke Göran Samuelsson 《Planta》1995,195(3):345-351
An intracellular carbonic anhydrase (CA; EC 4.2.1.1) was purified and characterised from the unicellular green alga Coccomyxa sp. Initial studies showed that cultured Coccomyxa cells contain an intracellular CA activity around 100 times higher than that measured in high-CO2-grown cells of Chlamydomonas reinhardtii CW 92. Purification of a protein extract containing the CA activity was carried out using ammonium-sulphate precipitation followed by anion-exchange chromatography. Proteins were then separated by native (non-dissociating) polyacrylamide gel electrophoresis, with each individual protein band excised and assayed for CA activity. Measurements revealed CA activity associated with two discrete protein bands with similar molecular masses of 80 +5 kDa. Dissociation by denaturing polyacrylamide gel electrophoresis showed that both proteins contained a single polypeptide of 26 kDa, suggesting that each 80-kDa native protein was a homogeneous trimer. Isoelectric focusing of the 80-kDa proteins also produced a single protein band at a pH of 6.5. Inhibition studies on the purified CA extract showed that 50% inhibition of CA activity was obtained using 1 M azetazolamide. Polyclonal antibodies against the 26-kDa CA were produced and shown to have a high specific binding to a single polypeptide in soluble protein extracts from Coccomyxa cells. The same antiserum, however, failed to cross-react with soluble proteins isolated from two different species of green algae, Chlamydomonas reinhardtii and Chlorella vulgaris. Correspondingly, antisera directed against pea chloroplastic CA, extracellular CA from C. reinhardtii and human CAII, showed no cross-hybridisation to the 26-kDa polypeptide in Coccomyxa. The 26-kDa protein was confirmed as being a CA by N-terminal sequencing of two internal polypeptide fragments and alignment of these sequences with that of previously identified CA proteins from several different species.Abbreviations CA
carbonic anhydrase
- CCM
CO2-concentrating mechanism
- IEF
isoelectric focusing
- Rubisco
ribulose-l,5-bisphosphate carboxylase/oxygenase
We would like to thank Drs. Cecilia Forsman, Inga-Maj Johansson and Nalle Jonsson for their valuable advice concerning the isolation of CA. This work was supported by the Swedish Natural Research Council and Seth M. Kempes Memorial foundation. 相似文献
8.
We examined cytoplasmic pH regulation inSchizosaccharomyces pombe andSaccharomyces cerevisiae using pH-sensitive fluorescent dyes. Of several different fluorescent compounds tested, carboxy-seminaphthorhodafluor-1 (C.SNARF-1) was the most effective. Leakage of C.SNARF-1 fromS. pombe was much slower than leakage fromC. cerevisiae. Using the pH-dependent fluorescence of C.SNARF-1 we showed that at an external pH of 7, mean resting internal pH was 7.0 forS. pombe and 6.6 forS. cerevisiae. We found that internal pH inS. pombe was maintained over a much narrower range in response to changes in external pH, especially at acidic pH. The addition of external glucose caused an intracellular alkalinization in both species, although the effect was much greater inS. cerevisiae than inS. pombe. The plasma membrane H+-ATPase inhibitor diethylstilbestrol reduced both the rate and extent of alkalinisation, with an IC50 of approximately 35 M in both species. Amiloride also inhibited internal alkalinisation with IC50's of 745 M forS. cerevisiae and 490 M forS. pombe.Abbreviations C.SNARF-1
carboxy-seminaphthorhodafluor-1 (-AM-acetoxy-methylester)
- DES
diethylstilbestrol
- IC50
apparent inhibitory constant
- BCECF
2,7-bis-(carboxyethyl)-5(6)-carboxyfluorescein (-AM--pentaacetoxymethyl ester)
- FDA
fluorescein diacetate 相似文献
9.
Johann S. Schwegler Wieland Steigner Almut Heuner Stefan Silbernagl 《The Journal of membrane biology》1990,117(3):243-251
Summary Confluent monolayers of the established opossum kidney cell line were exposed to NH4Cl pulses (20 mmol/liter) during continuous intracellular measurements of pH, membrane potential (PD
m
) and membrane resistance (R
m) in bicarbonate-free Ringer. The removal of extracellular NH4Cl leads to an intracellular acidification from a control value of 7.33±0.08 to 6.47±0.03 (n=7). This inhibits the absolute K conductance (g
K+), reflected by a decrease of K+ transference number from 71±3% (n=28) to 26±6% (n=5), a 2.6±0.2-fold rise ofR
m, and a depolarization by 24.2±1.5 mV (n=52). In contrast, intracellular acidification during a block ofg
K+ by 3 mmol/liter BaCl2 enhances the total membrane conductance, being shown byR
m decrease to 68±7% of control and cell membrane depolarization by 9.8±2.8 mV (n=17). Conversely, intracellular alkalinization under barium elevatesR
m and hyperpolarizes PD
m
. The replacement of extracellular sodium by choline in the presence of BaCl2 significantly hyperpolarizes PD
m
and increasesR
m, indicating the presence of a sodium conductance. This conductance is not inhibited by 10–4 mol/liter amiloride (n=7). Patch-clamp studies at the apical membrane (excised inside-out configuration) revealed two Na+-conductive channels with 18.8±1.4 pS (n=10) and 146 pS single-channel conductance. Both channels are inwardly rectifying and highly selective towards Cl–. The low-conductive channel is 4.8 times more permeable for Na+ than for K+. Its open probability rises at depolarizing potentials and is dependent on the pH of the membrane inside (higher at pH 6.5 than at pH 7.8). 相似文献
10.
The effects of phosphorus, Zn2+, CO2, and light intensity on growth, biochemical composition, and the activity of extracellular carbonic anhydrase (CA) in Isochrysis galbana were investigated. A significant change was observed when the concentration of phosphorus in the medium was increased from
5 μmol/L to 1000 μmol/L affecting I. galbana’s cell density, biochemical composition, and the activity of extracellular CA. Phosphorous concentration of 50 μmol/L to 500
μmol/L was optimal for this microalgae. The Zn2+ concentration at 10 μmol/L was essential to maintain optimal growth of the cells, but a higher concentration of Zn2+ (≥ 1000 μmol/L) inhibited the growth of I. galbana. High CO2 concentrations (43.75 mL/L) significantly increased the cell densities compared to low CO2 concentrations (0.35 mL/L). However, the activity of extracellular CA decreased significantly with an increasing concentration
of CO2. The activity of extracellular CA at a CO2 concentration of 43.75 mL/L was approximately 1/6 of the activity when the CO2 concentration was at 0.35 mL/L CO2. Light intensity from 4.0 mW/cm2 to 5.6 mW/cm2 was beneficial for the growth, biochemical composition and the activity of extracellular CA. The lower and higher light intensity
was restrictive for growth and changed its biochemical composition and the activity of extracellular CA. These results indicate
that phosphorus, Zn2+, CO2, and light intensity are important factors that impact growth, biochemical composition and the activity of extracellular
CA in I. galbana. 相似文献
11.
Kupriianova EV Markelova AG Lebedeva NV Gerasimenko LM Zavarzin GA Pronina NA 《Mikrobiologiia》2004,73(3):307-311
The activity of carbonic anhydrase (CA) was studied in different cell fractions of the alkaliphilic cyanobacterium Microcoleus chthonoplastes. The activity of this enzyme was found in the soluble and membrane protein fractions, as well as in intact cells and in a thick glycocalyx layer enclosing the cyanobacterium cells. The localization of CA in glycocalyx of M. chthonoplastes was shown by the western blot analysis and by immunoelectron microscopy studies with antibodies to the thylakoid CA from Chlamydomonas reinhardtii (Cah3). At least one of the CA forms occurring in M. chthonoplastes CA was shown to be an alpha-type enzyme. A possible mechanism of the involvement of the glycocalyx CA in calcification of cyanobacteria is discussed. 相似文献
12.
Summary
Dunaliella acidophila is an unicellular green alga which grows optimally at pH 0–1 while maintaining neutral internal pH. A plasma membrane preparation of this algae has been purified on sucrose density gradients. The preparation exhibits vanadatesensitive ATPase activity of 2 mol Pi/mg protein/min, an activity 15 to 30-fold higher than that in the related neutrophilic speciesD. salina. The following properties suggest that the ATPase is an electrogenic plasma membrane H+ pump. (i) ATP induces proton uptake and generates a positive-inside membrane potential as demonstrated with optical probes. (ii) ATP hydrolysis and proton uptake are inhibited by vanadate, diethylstilbestrol, dicyclohexylcarbodiimide and erythrosine but not by molybdate, azide or nitrate. (iii) ATP hydrolysis and proton uptake are stimulated by fussicoccin in a pH-dependent manner as found for plants plasma membrane H+-ATPase. Unusual properties of this enzyme are: (i) theK
m for ATP is around 60 M, considerably lower than in other plasma membrane H+-ATPases, and (ii) the ATPase activity and proton uptake are stimulated three to fourfold by K+ and to a smaller extent by other monovalent cations. These results suggest thatD. acidophila possesses a vanadate-sensitive H+-ATPase with unusual features enabling it to maintain the large transmembrane pH gradient. 相似文献
13.
D. A. Colin H. Aguenaou 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1992,162(3):219-223
Summary This paper presents a study of the mechanisms of Cl– transport through the brush border membranes of the posterior part of the intestine in the freshwater trout, Oncorhynchus mykiss. The mechanisms for Cl– transport in the posterior intestine are distinct from those in the middle intestine; an inwardly directed pH gradient stimulates Cl– uptake by bursh border membrane vesicles, indicating a Cl–/OH– exchange. A pH-regulated Cl– conductance is present, which is not activated at normal intracellular pH. Cl– uptake is stimulated by an outwardly directed HCO
3
–
gradient revealing the presence of a Cl–/HCO
3
–
exchange but, conversely, Cl– is not exchanged against SO
4
2-
. In addition, carbonic anhydrase activities have been detected in both the intracellular and extracellular leaflets of the bursh border membranes which favour the establishment of a bicarbonate gradient. A model of Cl– transport mechanisms through the brush-border membranes of the posterior intestine of the freshwater trout is proposed.Abbreviations BBM
brush border membrane
- CA
carbonic anhydrase
- EGTA
ethylene-bis(oxyethylenenitrilo)tetra-acetic acid
- FW
fresh water
- Hepes
N-2-hydroxy-ethyl-piperazine-N'-2-ethanesulphonic acid
- Mes
2-(N-morpholino)ethane sulphonic acid
- SITS
4-acetamido-4-isothiocyanostilbene-2,2-disulphonic acid
- TEA
triethanolamine
- TMA
tetramethylammonium
- TRIS
tris(hydroxymethyl)aminomethane 相似文献
14.
S. S. Madsen B. K. Larsen F. B. Jensen 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1996,166(2):101-109
Osmoregulation, acid-base balance and respiratory parameters were investigated in whitefish following transfer from freshwater to salt water. Whitefish acclimated successfully to 25 ppt brackish water but died after direct transfer to 32 ppt sea water. Transfer to brackish water induced rapid (<6 h) and permanent increases in plasma [Na+], [Cl–], total [Ca] and [Mg]. The extracellular hyperosmolality effected a transient (<3 days) muscle tissue dehydration and red blood cell shrinkage. Exposure to brackish water decreased both the arterial O2 tension and whole body O2 uptake. The extracellular acid-base status changed from an initial respiratory acidosis at 1 h towards a pronounced metabolic acidosis at 48 h of brackish water exposure. Red cell pHi decreased in parallel with extracellular pHe, but the in vivo pHi/pHe was only 0.26, suggesting some selective protection of red cell pHi. Plasma cortisol concentration and gill Na+, K+-ATPase activity increased after exposure to high ambient salinity, reflecting the induction of hypo-osmoregulatory mechanisms. The physiological changes in whitefish are discussed in relation to salinity-induced effects in other salmonid fishes.Abbreviations
CO2 solubility in plasma
-
water O2 capacitance coefficient
-
BW
brackish water
-
C
T
total CO2 content in plasma
-
FW
fresh water
-
Hb
hemoglobin
-
Hct
hematocrit
-
M
b
body mass of fish
-
MCHC
mean cellular hemoglobin concentration
-
PCO2
carbon dioxide tension
-
pH
e
extracellular pH
-
pH
i
intracellular pH
-
PO2 in
oxygen tension in water flowing in
-
PO2 out
oxygen tension in water flowing out
-
ppt
parts per thousand
-
RBC
red blood cell(s)
-
SW
sea water
-
V
m
water flows through chamber
-
OV
2
ml O2 consumed per kg per hour 相似文献
15.
Using a Ca2+-sensitive fluorescent indicator, fura-2/AM, we recorded calcium transients in secretory cells of isolated acini of the rat submandibular salivary gland; these transients were induced by hyperpotassium-induced depolarization (after an increase in [K+]
e
up to 50 mM) of the plasma membrane of the above cells. Calcium transients were significantly suppressed by 50 M nifedipine. Addition of 10 M carbonyl cyanide m-chlorophenylhydrazone to the normal extracellular solution was accompanied by a rise in [Ca2+]
i
, whereas when hyperpotassium solution is used the effect was less expressed. Blockers of CA2+-ATPase in the cellular membrane and in the endoplasmic reticulum, eosin Y (5 M) and cyclopiazonic acid (CPA, 5 M), respectively, evoked a significant increase in [Ca2+]
i
and a decrease in the K+-depolarization-induced calcium transient. Extracellular application of caffeine (2, 10, or 30 mM) was accompanied by a concentration-dependent rise in [Ca2+]
i
. Therefore, potassium depolarization of the plasma membrane of acinar cells of the rat submandibular salivary gland activates both the voltage-dependent Ca2+ influx and Ca2+-induced Ca2+ release from the endoplasmic reticulum; the initial level of [Ca2+]
i
was restored at the joint involvement of Ca2+-ATPases in the plasma membrane and the membranes of the endoplasmic reticulum and mitochondria. 相似文献
16.
Summary The rate of inorganic carbon uptake and its steadystate accumulation ratio (intracellular/extracellular concentration) was determined in the cyanobacteriumAnabaena variabilis as a function of extracellular pH. The free energy of protons (
) across the plasmalemma was calculated from determinations of membrane potential, and intracellular pH, as a function of the extracellular pH. While inward proton motive force decreased with increasing extracellular pH from 6.5 to 9.5, rate of HCO
3
–
influx and its accumulation ration increased. The latter is several times larger than would be expected should HCO
3
–
influx be driven by
. It is concluded that HCO
3
–
transport in cyanobacteria is not driven by the proton motive force. 相似文献
17.
Pronina N. A. Allakhverdiev S. I. Kupriyanova E. V. Klyachko-Gurvich G. L. Klimov V. V. 《Russian Journal of Plant Physiology》2002,49(3):303-310
The partitioning of carbonic anhydrase (CA) activity in chloroplasts isolated from 10–14-day-old pea (Pisum sativum L.) seedlings was investigated. The effect of CA inhibitors on the kinetics of chlorophyll fluorescence in photosystem II (PSII) preparations was also studied. The activity of CA was detected in fractions of soluble proteins and in the polypeptide complexes of the PSI and PSII. Isolated particles of photosystems retained a high photochemical activity similar to that of intact chloroplasts and the high level of polyunsaturated fatty acids. The association of CA with the particles of PSII (PSII-CA) was also tested by Western-blot analysis using antibodies against PSII-CA (Cah3) from Chlamydomonas reinhardtii. The PSII particles isolated with Triton X-100 (T-20) showed a higher activity of the enzyme as calculated on a protein basis than the DT-20 particles isolated with digitonin and Triton X-100. This difference seems to be related to the higher degree of nativity of the chloroplast T-20 fragments as compared to DT-20 particles. The higher level of chlorophyll per reaction center as well as the higher content of chlorophyll b and lipid fatty acids as calculated on protein basis, in particular of E-16:113 acid, which stabilizes the oligomeric structure of the light-harvesting complex of the PSII, also confirms this suggestion. The activity of CA was not detected in the DT-20 preparations treated with Tris–HCl to eliminate manganese ions. This is likely to indicate that one of the extrinsic polypeptides of PSII exhibits CA activity. Specific inhibitors of CA (acetazolamide and imidazole) inhibited the photoinduced yield of chlorophyll fluorescence (F). This might be determined by damaging the water-oxidizing system or its interaction with the PSII reaction centers. The functional role of PSII-CA for 2-concentrating in carboxylation sites as well as its role in the coupling of light and dark reactions in chloroplasts is discussed. 相似文献
18.
J. J. H. H. M. De Pont S. E. Van Emst-De Vries S. L. Bonting 《Journal of bioenergetics and biomembranes》1984,16(4):263-281
The effects of three amino group reagents on the activity of (Na++K+)-ATPase3 and its component K+-stimulatedp-nitrophenylphosphatase activity from rabbit kidney outer medulla have been studied. All three reagents cause inactivation of the enzyme. Modification of amino groups with trinitrobenzene sulfonic acid yields kinetics of inactivation of both activities, which depend on the type and concentration of the ligands present. In the absence of added ligands, or with either Na+ of Mg2+ present, the enzyme inactivation process follows complicated kinetics. In the presence of K+, Rb+, or Tl+, protection occurs due to a change of the kinetics of inactivation toward a first-order process. ATP protects against inactivation at a much lower concentration in the absence than in the presence of Mg2+ (P
50 6 µM vs. 1.2 mM). Under certain conditions (100 µM reagent, 0.2 M triethanolamine buffer, pH 8.5) modification of only 2% of the amino groups is sufficient to obtain 50% inhibition of the ATPase activity. Modification of amino groups with ethylacetimidate causes a nonspecific type of inactivation of (Na++K+)-ATPase. Mg2+ and K+ have no effects, and ATP only a minor effect, on the degree of modification. The K+-stimulatedp-nitrophenylphosphatase activity is less inhibited than the (Na++K+)-ATPase activity. Half-inhibition of the (Na++K+)-ATPase is obtained only after 25% modification of the amino groups. Modification of amino groups with acetic anhydride also causes nonspecific inactivation of (Na++K+)-ATPase. Mg2+ has no effect, and ATP has only a slight protecting effect. The K+-stimulatedp-nitrophenylphosphatase activity is inhibited in parallel with the (Na++K+)-ATPase activity. Half-inactivation of the (Na++K+)-ATPase activity is obtained after 20% modification of the amino groups.This article is No. 52 in the series Studies on (Na++K+)-Activated ATPase. 相似文献
19.
Hubert Felle 《Planta》1988,176(2):248-255
In cells of Zea mays (root hairs, coleoptiles) and Riccia fluitans (rhizoids, thalli) intracellular Ca2+ and pH have been measured with double-barrelled microelectrodes. Free Ca2+ activities of 109–187 nM (Riccia rhizoids), 94–160 nM (Riccia thalli), 145–231 nM (Zea root hairs), 84–143 nM (Zea coleoptiles) were found, and therefore identified as cytoplasmic. In a few cases (Riccia rhizoids), free Ca2+ was in the lower millimolar range (2.3±0.8 mM). A change in external Ca2+ from 0.1 to 10 mM caused an initial and short transient increase in cytoplasmic free Ca2+ which finally levelled off at about 0.2 pCa unit below the control, whereas in the presence of cyanide the Ca2+ activity returned to the control level. It is suggested that this behaviour is indicative of active cellular Ca2+ regulation, and since it is energy-dependent, may involve a Ca2+-ATPase. Acidification of the cytoplasmic pH and alkalinization of the vacuolar pH lead to a simultaneous increase in cytoplasmic free Ca2+, while alkalinization of pHc decreased the Ca2+ activity. Since this is true for such remote organisms as Riccia and Zea, it may be concluded that regulation of cytoplasmic pH and free Ca2+ are interrelated. It is further concluded that double-barrelled microelectrodes are useful tools for investigations of intracellular ion activities in plant cells.Symbols and abbreviations m, m
membrane potential difference, changes thereof
- PVC
polyvinylchloride 相似文献
20.
Na+/H+ antiporters are universal devices involved in the Na+ and H+ circulation of both eukaroyotes and prokaryotes, thus playing an essential role in the pH and Na+ homeostasis of cells. This review focuses on the major impact of the application of molecular biology tools in the study of the antiporters. These tools permit the verification of the role of the antiporters and provide insights into their unique biology. A novel signal transduction to Na+ involvingnhaR, a positive regulator, controls the expression ofnhaA inE. coli. A pH sensor regulates the activity of Na+/H+ antiporters, both in eukaryotes and prokaryotes. A most intricate signal transduction to pH involving phosphorylation steps controls the activity ofnhel in higher mammals. The identification of Histidine 226 in the pH sensor of NhaA is a step forward towards the understanding of the pH regulation of these proteins. 相似文献