共查询到20条相似文献,搜索用时 15 毫秒
1.
Carbonic anhydrase (CA) activity of pea thylakoids, thylakoid membranes enriched with photosystem I (PSI-membranes), or photosystem
II (PSII-membranes) as well as both supernatant and pellet after precipitation of thylakoids treated with detergent Triton
X-100 were studied. CA activity of thylakoids in the presence of varying concentrations of Triton X-100 had two maxima, at
Triton/chlorophyll (triton/Chl) ratios of 0.3 and 1.0. CA activities of PSI-membranes and PSII-membranes had only one maximum
each, at Triton/Chl ratio 0.3 or 1.0, respectively. Two CAs with characteristics of the membrane-bound proteins and one CA
with characteristics of the soluble proteins were found in the medium after thylakoids were incubated with Triton. One of
the first two CAs had mobility in PAAG after native electrophoresis the same as that of CA residing in PSI-membranes, and
the other CA had mobility the same as the mobility of CA residing in PSII-membranes, but the latter was different from CA
situated in PSII core-complex (Ignatova et al. 2006 Biochemistry (Moscow) 71:525–532). The properties of the “soluble” CA removed from thylakoids were different from the properties
of the known soluble CAs of plant cell: apparent molecular mass was about 262 kD and it was three orders more sensitive to
the specific CA inhibitor, ethoxyzolamide, than soluble stromal CA. The data are discussed as indicating the presence of,
at least, four CAs in pea thylakoids. 相似文献
2.
Gamble W 《Journal of theoretical biology》2006,239(1):16-21
Atherosclerosis is a dynamic multifaceted disease which affects the aorta and its major branches, characterized by the presence of lesions called atheromatous plaques. The plaque is a focal thickening of the intima caused by proliferation of smooth muscle cells, and the deposition of cholesterol, other lipids, hydroxyapatite and fibrous connective tissue. It is proposed that the determinant step of the process which leads to the disease atherosclerosis is the calcium precipitation which traps cholesterol in the plaque precursor matrix which contains lipoproteins, calcium carbonate, hydroxyapapatite, triglycerides, albumin, calmodulin and other proteins. The bear, a species which does not contract the disease is used as an example in support of the hypothesis. The bear's ability to regulate calcium levels and the regulation of acid base balance via regulation of carbon dioxide levels permits the control of the determinant step of plaque formation, that is calcification of the plaque. 相似文献
3.
Mikulski R Domsic JF Ling G Tu C Robbins AH Silverman DN McKenna R 《Archives of biochemistry and biophysics》2011,516(2):97-102
The tryptophan residue Trp5, highly conserved in the α class of carbonic anhydrases including human carbonic anhydrase II (HCA II), is positioned at the entrance of the active site cavity and forms a π-stacking interaction with the imidazole ring of the proton shuttle His64 in its outward orientation. We have observed that replacement of Trp5 in HCA II caused significant structural changes, as determined by X-ray diffraction, in the conformation of 11 residues at the N-terminus and in the orientation of the proton shuttle residue His64. Most significantly, two variants W5H and W5E HCA II had His64 predominantly outward in orientation, while W5F and wild type showed the superposition of both outward and inward orientations in crystal structures. Although Trp5 influences the orientation of the proton shuttle His64, this orientation had no significant effect on the rate constant for proton transfer near 1 μs−1, determined by exchange of 18O between CO2 and water measured by mass spectrometry. The apparent values of the pKa of the zinc-bound water and the proton shuttle residue suggest that different active-site conformations influence the two stages of catalysis, the proton transfer stage and the interconversion of CO2 and bicarbonate. 相似文献
4.
Demian Riccardi Shuo Yang Qiang Cui 《Biochimica et Biophysica Acta - Proteins and Proteomics》2010,1804(2):342-351
Recent QM/MM analyses of proton transfer function of human carbonic anhydrase II (CAII) are briefly reviewed. The topics include a preliminary analysis of nuclear quadrupole coupling constant calculations for the zinc ion and more detailed analyses of microscopic pKa of the zinc-bound water and free energy profile for the proton transfer. From a methodological perspective, our results emphasize that performing sufficient sampling is essential to the calculation of all these quantities, which reflects the well solvated nature of CAII active site. From a mechanistic perspective, our analyses highlight the importance of electrostatics in shaping the energetics and kinetics of proton transfer in CAII for its function. We argue that once the pKa for the zinc-bound water is modulated to be in the proper range (~ 7.0), proton transfer through a relatively well solvated cavity towards/from the protein surface (His64) does not require any major acceleration. Therefore, although structural details like the length of the water wire between the donor and acceptor groups still may make a non-negligible contribution, our computational results and the framework of analysis suggest that the significance of such “fine-tuning” is likely secondary to the modulation of pKa of the zinc-bound water. We encourage further experimental analysis with mutation of (charged) residues not in the immediate neighborhood of the zinc ion to quantitatively test this electrostatics based framework; in particular, Φ analysis based on these mutations may shed further light into the relative importance of the classical Grotthus mechanism and the “proton hole” pathway that we have proposed recently for CAII. 相似文献
5.
6.
Anjana Sharma Abhishek Bhattacharya Rajesh Pujari Ankita Shrivastava 《Indian journal of microbiology》2008,48(3):365-371
Diversified group of bacteria were screened for carbonic anhydrase (CA) activity. Significant CA activity was found in crude
enzyme extracts of Enterobacter and Aeromonas isolates while minimal or negligible CA activity was observed in case of Shigella and Klebsiella spp. Optimization and characterization study of potent CA producing isolates revealed that the maximum enzyme activity of
3.86 EU/ml was observed in E. taylorae and the optimum pH range for enzyme stability was found to be 7.5–9.0 along with an optimum temperature range of 35–50 °C.
The molecular mass of CA was 29-kDa indicating α-type with periplasmic and cytosolic location. Present investigation for the
first time reports CA in diversified genus and optimized parameters for enhanced production of CA in Enterobacter sp. & Aeromonas sp. from fresh water bodies that inturn lay down grounds for exploitation of CA from E. taylorae as an efficient catalyst for CO2 sequestration within a bioreactor. 相似文献
7.
Christine A. Raines Philip R. Horsnell Caroline Holder Julie C. Lloyd 《Plant molecular biology》1992,20(6):1143-1148
A full-length cDNA clone encoding carbonic anhydrase was isolated from an Arabidopsis thaliana (Columbia) leaf library. Comparison of the derived amino acid sequence obtained from this clone with those of pea and spinach reveals a considerable degree of identity. The carbonic anhydrase cDNA was used to probe the level of RNA encoding this protein in the leaves of plants grown in elevated CO2 (660 ppm). We have found that under these conditions the steady-state level of carbonic anhydrase mRNA was increased in comparison with control plants grown in normal atmospheric concentrations of CO2 (330 ppm). This raises the intruiging possibility that there exists in higher plants a mechanism for perceiving and responding to changes in environmental CO2 concentrations at the genetic level. 相似文献
8.
The review summarizes current data on the existence in terrestrial higher plants of several carbonic anhydrase forms differing in their properties, molecular structure, and intracellular localization. Possible functions of these carbonic anhydrases are discussed as well as specific features of carbon-concentrating mechanisms in phototrophic tissues of plants with C3 and C4 pathways of photosynthesis. 相似文献
9.
Secil Deniz Tugba Kevser Uysal Clemente Capasso Claudiu T. Supuran Ozen Ozensoy Guler 《Journal of enzyme inhibition and medicinal chemistry》2021,36(1):1230
The ongoing Covid-19 is a contagious disease, and it is characterised by different symptoms such as fever, cough, and shortness of breath. Rising concerns about Covid-19 have severely affected the healthcare system in all countries as the Covid-19 outbreak has developed at a rapid rate all around the globe. Intriguing, a clinically used drug, acetazolamide (a specific inhibitor of carbonic anhydrase, CA, EC 4.2.1.1), is used to treat high-altitude pulmonary oedema (HAPE), showing a high degree of clinical similarities with the pulmonary disease caused by Covid-19. In this context, this preliminary study aims to provide insights into some factors affecting the Covid-19 patients, such as hypoxaemia, hypoxia as well as the blood CA activity. We hypothesise that patients with Covid-19 problems could show a dysregulated acid–base status influenced by CA activity. These preliminary results suggest that the use of CA inhibitors as a pharmacological treatment for Covid-19 may be beneficial. 相似文献
10.
AbstractCarbonic anhydrase (CA) is the most effective CO2 hydratase catalyst, but the poor storage stability and repeatability of CA limit its development. Therefore, CA was immobilized on the epoxy magnetic composite microspheres to enhance the CO2 absorption into N-methyldiethanolamine (MDEA) aqueous solution in this work. In the presence of immobilized CA, the CO2 absorption rate of MDEA solution (10?wt%) (0.63?mmol·min?1) was greatly improved by almost 40%, and their reaction equilibrium time was shortened from 150?min to 90?min compared with that into MDEA solution. The results indicated that the absorption of CO2 into MDEA solution had been significantly enhanced by using CA. After the 7th reuse recycle, the activity of the immobilized CA was still closed to its initial value at 313.15?K. Moreover, enzyme catalytic kinetics of immobilized CA was investigated using the p-nitrophenyl acetate (p-NPA) as substrate. The values of Michaelis–Menten constant (Km) and the maximum velocity (Vmax) of the immobilized CA were calculated to be 27.61?mmol/L and 20.14?×?10?3?mmol·min?1·mL?1, respectively. Besides, the kinetics of CO2 reaction into MDEA with or without CA were also compared. The results showed that CO2 absorption into CA/MDEA aqueous solution obeyed the pseudo first order regime and the second order kinetics rate constant (k2) was calculated to be 929?m3·kmol?1·s?1, which was twice higher than that of MDEA aqueous solution without immobilized CA (k2=414 m3·kmol?1·s?1) at 313.15?K. 相似文献
11.
AbstractThe novel technology of biological carbon sequestration using microbial enzymes have numerous advantages over conventional sequestration strategies. In the present study, extracellular carbonic anhydrase (CA) producing bacteria were isolated from water samples in the Arabian Sea, India. A potential isolate, Bacillus safensis isolate AS-75 was identified based on 16S rDNA sequence analysis. The culture conditions suitable for CA production were 32?°C incubation temperature with 4% NaCl and 10?mM Zn supplementation. Experimental optimization of culture conditions enhanced enzyme activity to 265?U mL?1. CA specific gene was characterized and based on the analysis, the CA of B. safensis isolate AS-75 was a leucine (11.3%) with α-helices as the dominant component in its secondary structure. Based on soil microcosm studies, CA could sequester CO2 by 95.4%?±?0.11% in sterilized soil with enzyme microcosm. Hence, the application of enzyme was found to be more effective in removing CO2. 相似文献
12.
The metabolic pathway of primary carbon fixation was studied in a peculiar pennate marine diatom, Haslea ostrearia (Bory) Simonsen, which synthesizes and accumulates a blue pigment known as “marennine”. Cells were cultured in a semi-continuous
mode under saturating [350 μmol(photon) m−2 s−1] or non-saturating [25 μmol(photon) m−2 s−1] irradiance producing “blue” (BC) and “green” (GC) cells, characterized by high and low marennine accumulation, respectively.
Growth, pigment contents (chlorophyll a and marennine), 14C accumulation in the metabolites, and the carbonic anhydrase (CA) activity of the cells were determined during the exponential
growth phase. Growth rate and marennine content were closely linked to irradiance during growth: higher irradiance increased
both growth rate and marennine content. On the other hand, the Chl a concentration was lower under saturating irradiance. The distribution between the Calvin-Benson (C3) and β-carboxylation (C4) pathways was very different depending on the irradiance during growth. Metabolites of the C3 cycle contained about 70 % of the total fixed radioactivity after 60 s of incorporation into cells cultured under the non-saturating
irradiance (GC), but only 47 % under saturating irradiance (BC). At the same time, carbon fixation by β-carboxylation was
24 % in GC versus about 41 % in BC, becoming equal to that in the C3 fixation pathway in the latter. Internal CA activity remained constant, but the periplasmic CA activity was higher under
low than high irradiance. 相似文献
13.
The activity of enzyme carbonic anhydrase (CA) was investigated in two diazotrophic cyanobacteria, Anabaena sp. (ARM 629) and Nostoc calcicola, in the presence of CO2/NaHCO3 and different inhibitors. The CA activity increased when the cells were pretreated with a high concentration of CO2/NaHCO3 and then transferred to ambient level CO2. Maximum activity of CA was observed after 8 h of incubation in light on transfer of cells from high Ci to ambient level CO2, and was low when incubated in dark. Addition of the photosynthetic inhibitor DCMU brought about a differential reduction in CA activity, depending on the carbon source (NaHCO3/CO2). CA inhibitors--ethoxyzolamide (EZ) and acetazolamide (AZ)--inhibited the enzyme activity in both the genera, but the extent of inhibition was greater in Anabaena sp. than in N. calcicola. Such a variation in extent of inhibition/stimulation of CA activity being different in the two genera reflects differences in their inherent potential and genetic background. The relevance of such cyanobacterial strains as CO2 sinks is also discussed. 相似文献
14.
The organization of carbonic anhydrase (CA) system in halo- and alkaliphilic cyanobacterium Rhabdoderma lineare was studied by Western blot analysis and immunocytochemical electron microscopy. The presence of putative extracellular α-CA of 60 kDa in the glycocalyx, forming a tight sheath around the cell, and of two intracellular β-CA is reported. We show for the first time that the β-CA of 60 kDa is expressed constitutively and associated with polypeptides of photosystem II (β-CA-PS II). Another soluble β-CA of 25 kDa was induced in low-bicarbonate medium. Induction of synthesis of the latter β-CA was accompanied by an increase in the intracellular pool of inorganic carbon, which suggests an important role of this enzyme in the functioning of a CO2-concentrating mechanism. 相似文献
15.
An Integrated Vacuum Carbonate Absorption Process (IVCAP) currently under development could significantly reduce the energy consumed when capturing CO2 from the flue gases of coal-fired power plants. The biocatalyst carbonic anhydrase (CA) has been found to effectively promote the absorption of CO2 into the potassium carbonate solution that would be used in the IVCAP. Two CA enzymes were immobilized onto three selected support materials having different pore structures. The thermal stability of the immobilized CA enzymes was significantly greater than their free counterparts. For example, the immobilized enzymes retained at least 60% of their initial activities after 90 days at 50 °C compared to about 30% for their free counterparts under the same conditions. The immobilized CA also had significantly improved resistance to concentrations of sulfate (0.4 M), nitrate (0.05 M) and chloride (0.3 M) typically found in flue gas scrubbing liquids than their free counterparts. 相似文献
16.
A photoacoustic pulse-modulation technique is applied for the study of a CO2-stimulated gas uptake signal in leaves (Reising and Schreiber, Photosynthe Res 31: 227–238, 1992). It is shown that this uptake signal can be substantially suppressed by application of the carbonic anhydrase inhibitor, ethoxyzolamide, to leaf discs. This inhibitor does not affect the O2-evolution signal in air or the chlorophyll fluorescence induction pattern at high CO2, when non-saturating light intensities are used. On the basis of these findings it is concluded that at least a major part of the CO2-stimulated photoacoustic uptake signal results from light-modulated CO2-solubilisation catalysed by carbonic anhydrase. Modulated CO2-solubilisation appears likely to be induced by light driven H+-translocation from the stroma into the thylakoid lumen. Comparison of the induction patterns of chlorophyll fluorescence quenching and the uptake signal suggests a correlation between membrane energisation and CO2-uptake. The importance of O2-dependent electron flow as a major cause of membrane energisation is discussed. It is proposed that in the absence of CO2 the combination of Mehler- and ascorbate peroxidase reactions does not result in a photobaric signal, as O2-uptake and O2-evolution components cancel each other. Two main conclusions, which are of considerable importance for future practical applications of the photoacoustic method, are drawn from these findings: (1) When high CO2 is applied to leaves, the photobaric uptake component may provide a unique means of monitoring the function of stromal carbonic anhydrase in vivo. (2) Brief flushing of the photoacoustic cell with air may prevent the occurrence of an uptake signal, thus allowing a straight-forward deconvolution into photothermal and O2-evolution components. 相似文献
17.
Klier M Schüler C Halestrap AP Sly WS Deitmer JW Becker HM 《The Journal of biological chemistry》2011,286(31):27781-27791
The ubiquitous enzyme carbonic anhydrase isoform II (CAII) has been shown to enhance transport activity of the proton-coupled monocarboxylate transporters MCT1 and MCT4 in a non-catalytic manner. In this study, we investigated the role of cytosolic CAII and of the extracellular, membrane-bound CA isoform IV (CAIV) on the lactate transport activity of the high-affinity monocarboxylate transporter MCT2, heterologously expressed in Xenopus oocytes. In contrast to MCT1 and MCT4, transport activity of MCT2 was not altered by CAII. However, coexpression of CAIV with MCT2 resulted in a significant increase in MCT2 transport activity when the transporter was coexpressed with its associated ancillary protein GP70 (embigin). The CAIV-mediated augmentation of MCT2 activity was independent of the catalytic activity of the enzyme, as application of the CA-inhibitor ethoxyzolamide or coexpressing the catalytically inactive mutant CAIV-V165Y did not suppress CAIV-mediated augmentation of MCT2 transport activity. Furthermore, exchange of His-88, mediating an intramolecular H(+)-shuttle in CAIV, to alanine resulted only in a slight decrease in CAIV-mediated augmentation of MCT2 activity. The data suggest that extracellular membrane-bound CAIV, but not cytosolic CAII, augments transport activity of MCT2 in a non-catalytic manner, possibly by facilitating a proton pathway other than His-88. 相似文献
18.
Wu Yanyou Li Xiteng Li Pingping Zhao Xinzheng 《Plant Cell, Tissue and Organ Culture》2006,84(1):124-127
During plant tissue culture, the culture container is small and sealed; the concentration of CO2 in the microenvironment is relatively low. The plantlet growth is restrained for the shortage of CO2 in the culture container. Carbonic anhydrase is a zinc-containing metalloenzyme that catalyzes the reversible conversion of bicarbonate to CO2. The determination of carbonic anhydrase of leaves from Atractylodes lancea (thunb.) DC, Orychophragmus violaceus (L.) O.E. Schulz, Brassica juncea (L.) Czern.et Coss. cv. Luzhousileng, Brassica campestris L. cv. Chuanyou No.8, Brassica napus L cv. Oro, Brassica carinata Braun, Raphanus sativa L. var. raphanistroides Makino and their plantlets indicates that the carbonic anhydrase activity of leaves from both plantlets and fields varies from plant species to plant species, the carbonic anhydrase activity of leaves of Atractylodes lancea (thunb.) DC is the lowest among those plants, and the leaves of all plantlets are lower in carbonic anhydrase activity than the same species of plants from fields. The comparison of the growth rates of those plantlets shows that their relative growth rates are significantly different, plantlets of Atractylodes lancea have the slowest relative growth rate among those plants, and plantlets of Brassica juncea have the greatest relative growth rate. The relationship between RGR of plantlets and their CA activities is a significant linear function. It seems that there was certain correlation between carbonic anhydrase activities of plants and their growth rates. It suggests that in vitro, the greater the carbonic anhydrase activity of plantlet is, the higher its net photosynthetic rate, and the faster its growth rate. Those results offer a foundation to a rational medium choice in plant tissue culture. 相似文献
19.
A cDNA encoding the mature, chloroplast-localized carbonic anhydrase in pea has been expressed inE. coli. The enzyme is fully active and yields of up to 20% of the total soluble protein can be obtained from the bacteria. This expression system was used to monitor the effects of site-directed mutagenesis of seven residues found within conserved regions in the pea carbonic anhydrase amino acid sequence. The effects of these modifications are discussed with respect to the potential of various amino acids to act as sites for zinc coordination or intramolecular proton shuttles. 相似文献
20.
Yong Yu Zhang Kathy E. Sietsema Cynthia S. Sullivan Karhnan Wasserman 《European journal of applied physiology and occupational physiology》1994,69(4):309-315
A method to estimate the CO2 derived from buffering lactic acid by HCO3
– during constant work rate exercise is described. It utilizes the simultaneous continuous measurement of O2 uptake (
O2) and CO2 output (
CO2), and the muscle respiratory quotient (RQm). The CO2 generated from aerobic metabolism of the contracting skeletal muscles was estimated from the product of the exercise-induced increase in
O2 and RQm calculated from gas exchange. By starting exercise from unloaded cycling, the increase in CO2 stores, not accompanied by a simultaneous decrease in O2 stores, was minimized. The total CO2 and aerobic CO2 outputs and, by difference, the millimoles (mmol) of lactate buffered by HCO3
– (corrected for hyperventilation) were estimated. To test this method, ten normal subjects performed cycling exercise at each of two work rates for 6 min, one below the lactic acidosis threshold (LAT) (50 W for all subjects), and the other above the LAT, midway between LAT and peak
O2 [mean (SD), 144 (48) W]. Hyperventilation had a small effect on the calculation of mmol lactate buffered by HCO3
– [6.5 (2.3)% at 6 min in four subjects who hyperventilated]. The mmol of buffer CO2 at 6 min of exercise was highly correlated (r = 0.925, P < 0.001) with the increase in venous blood lactate sampled 2 min into recovery (coefficient of variation = ±0.9 mmol·l–1). The reproducibility between tests done on different days was good. We conclude that the rate of release of CO22 from HCO3
– can be estimated from the continuous analysis of simultaneously measured
CO2,
O2, and an estimate of muscle substrate. 相似文献