The Spirulina platensis Adenylate Cyclase Gene, cyaC, Encodes a Novel Signal Transduction Protein (original) (raw)
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Properties of the cyanobacterial coupling factor ATPase from Spirulina platensis
Archives of Biochemistry and Biophysics, 1986
photosynthetic membranes and isolated F1 ATPase were characterized with respect to ATP activity. The following results indicate that the regulation of expression of ATPase activity in Spirulina platensis is similar to that found in chloroplasts: (a) the ATPase activity of Spirulina membranes and isolated Fi ATPase is mostly latent, a characteristic of ehloroplast ATPase activity; (b) treatments that elicit ATPase activity in higher plant chloroplast thylakoids and isolated chloroplast coupling factor (CF,) greatly stimulate the activity of Spirulina membranes and F1, and (c) the cation specificity of chloroplast ATPase activity, e.g., light-induced membrane activity that is magnesium dependent and trypsin-activated CFI activity that is calcium dependent, is also observed in Spirulina. Thus, an 8-to l&fold increase in specific activity (to 13-15 pm01 Pi min-' mg chl-') is obtained when Spirulina membranes are treated with trypsin (CaATPase) or with methanol (MgATPase); a lightinduced, dithiothreitol-dependent MgATPase activity is also found in the membranes. Purified Spirulinu F1 is a CaATPase when activated with trypsin (endogenous activity increases from 4 to 2'7-37 pmol Pi min-' mg protein-') or with dithiothreitol (5.6 hmol Pi min-' mg-l), but a MgATPase when assayed with methanol (18-20 pmol Pi min-' mg-'). The effects of varying calcium and ATP concentrations on the kinetics of trypsininduced CaATPase activity of Spirulina F1 were examined. When the calcium concentration is varied at constant ATP concentration, the velocity plot shows a marked sigmoidicity. By varying Ca-ATP metal-nucleotide complex concentration at constant concentrations of free calcium or ATP, it is shown that the sigmoidicity is due to the effect of free ATP, which changes the Hill constant to 1.6 from 1.0 observed when the free calcium concentration is kept constant at 5 mM. Therefore not only is ATP an inhibitor but it is also an allosteric effector of Spirulina F1 ATPase activity. At 5 MM free calcium, the Km for the Ca-ATP metal-nucleotide complex is 0.42 mM. o 19% Academic press, lnc.
Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1990
An adenylate cydase activity was partially characterized in the cyanobacterium Anabaena sp. The enzyme activity is found in soluble cell fractions and shows an apparent molecular weight of about 183400. This adenylate cyclase is activated by Ca 2+ and bovine brain or spinach calmodulin and it is inhibited by EGTA and some phenothiazine derivatives. Furthermore, Anabaena sp. extracts contain a calmodulin-like activity which stimulates bovine brain cyclic AMP phosphodiesterase and the A nabaena adenylate cyclase. EGTA and phenothiazine derivatives block the cyanobacterial modulator effect.
Phycocyanin promoter of Spirulina platensis controlling heterologous expression in cyanobacteria
Journal of Applied Phycology, 2011
An efficient expression vector for transferring exogenous deoxyribonucleic acid (DNA) into cell is important for biological study and recombinant protein production in cyanobacteria. The promoter, which can mediate gene expression and function in both Escherichia coli and cyanobacteria, could facilitate development of an expression vector for a DNA transfer system in cyanobacteria. Computational analysis of the phycocyanin (PC) promoter of Spirulina platensis strain C1 showed a high degree of conservation with the PC promoter of other cyanobacteria. In addition, the S. platensis PC promoter matched well with the consensus sequences of the E. coli promoter. Using green fluorescent protein (GFP) as a reporter, activity of the PC promoter of S. platensis C1 was functionally analyzed in Synechococcus elongatus and E. coli. The result revealed that the PC promoter could efficiently drive gfp expression in heterologous hosts, indicating the existence of common transcription machinery in these microbes. This promoter may be applicable not only for fundamental research but also for biotechnological purposes.
Photosynthetic phosphorylation by a membrane preparation of the cyanobacterium Spirulina Maxima
Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1982
A new method for the isolation of photosynthetic membranes from the cyanobacterium Spirulina maxima has been developed. When illuminated, these membranes evolve oxygen in the presence of ferricyanide (Hill reaction) and consume oxygen in the presence of methyl viologen (Mehler reaction). When the membranes are left to stand at 4°C for 30 min, they develop the ability to consume oxygen in the light without an added, artificial electron acceptor. The Hill and Mehler reactions are not affected by the presence of ADP or uncouplers, but are inhibited by triphenyitin chloride. We have detected a cryptic ATPase activity stimulated by trypsin in the 20000×g supernatant fraction of the membrane preparation. In addition, the membrane vesicles contain an ATPase activity which is enhanced by treatment with dithiothreitol in the presence of light. These observations of ATPase led us to try a careful titration of the membrane vesicles with both triphenyltin chloride and N,N'-dicyclohexylcarbodiimide. When the vesicles were sealed with these reagents, we could observe both cyclic and stoichiometric photosynthetic phosphorylation.
Biologia, 2020
Cyanobacteria are the only prokaryotes that can utilize energy sources from sunlight, electrons from water, carbon from the air, and have the capability of fixing nitrogen. The characterization of various cyanobacteria reveals the performance as well as growth differences when compared among different genus of cyanobacteria, which emphasizes the value of characterization in the cellular grounds. In this paper, we have performed a comparative study on the morphology, growth pattern, physiochemical properties as well as structural features of Spirulina sp. NCIM 5143 and Nostoc ellipsosporum NCIM 2786. The comparison is performed through microscopic images, FESEM and statistical study of cell growth by ANOVA. In silico approach was also carried out on the two enzymes present in both the cyanobacteria, Cyanophycin synthestase (EC: 6.3.2.29) and Nitrate reductase (EC: 1.7.1.1) by studying the physicochemical properties, sequential features and structural modeling. A descriptive profile study was completed on these microorganisms and their proteins that may help to interpret the molecular mechanism of the enzymatic reaction of 1, 3-Propanediol (PDO) production in the future.
Molecular Biology Reports, 2000
The genes from a cyanobacterium -Spirulina platensis strain C1 -that encode the acyl-lipid desaturases (desC, desA and desD) involved in γ -linolenic (GLA) synthesis have been successfully expressed for the first time in Escherichia coli by employing a pTrcHisA expression system. In this report, the authors describe the expression of the three Spirulina N-terminal 6xHis-desaturases as well as the functional analysis of these recombinant proteins. The gene products of desC, desA and desD have approximate molecular masses of 37, 45, and 47 kDa, respectively. Enzymatic activity measurement of these products was carried out in vivo to demonstrate that (i) the expressed proteins are in functional form, and (ii) the cofactors of the host system can complement the system of Spirulina platensis. The study demonstrated that the gene products of desC and desA catalyzed the reactions in vivo where the enzyme substrates were provided in appropriate concentration. This indicates that the 9 and 12 desaturases were expressed in the heterologous host in their active form, and that these two reactions can be carried out in an E. coli host cell using its cofactors system. In contrast, 6 desaturase activity can be detected only in vitro where electron carriers are provided. This suggests that while this enzyme is expressed in the heterologous host in its active form, its function in vivo is suppressed, as the electron carriers of the host system cannot complement the system of Spirulina platensis.
Molecular Biotechnology, 2007
Spirulina is distinguished from other cyanobacteria by its spiral morphology; however, this cyanobacterium has frequently been observed with a linear morphology in laboratory and industrial conditions. In our laboratory conditions, the simultaneously presence of the linear and spiral forms has also been observed. In the present study, the two forms of S. platensis C1 were separated and grown as axenic cultures in order to study the proteins that were differentially expressed in the soluble and insoluble protein fractions of the spiral and the linear forms. Two dimensional-differential gel electrophoresis (2D-DIGE) was performed to separate differentially expressed proteins that were subsequently identified by mass spectrometry. The differentially expressed proteins suggested two points. First, the morphological change is possibly induced by various environmental stresses such as oxygen level, carbon dioxide level, nutrient availability, and light. Second, the change of cell-shape might be a result of the change in a cell shape determination mechanism. Thus, this study is the first to show evidence at the protein level that may explain this morphological transformation in Spirulina.
MGG Molecular & General Genetics, 1989
A 5.3 kb DNA segment containing the str operon (ca. 4.5 kb) of the cyanobacterium Spirulina platensis has been sequenced. The str operon includes the structural genes rpsL (ribosomal protein S12), rpsG (ribosomal protein 7),fus(translationelongationfactorEF−G)andtuf(translationelongationfactorEF−Tu).Fromthenucleotidesequenceofthisoperon,theprimarystructuresofthefourgeneproductshavebeenderivedandcomparedwiththeavailablecorrespondingstructuresfromeubacteria,archaebacteriaandchloroplasts.Extensivehomologieswerefoundinalmostallcasesandintheorder7), fus (translation elongation factor EF-G) and tuf (translation elongation factor EF-Tu). From the nucleotide sequence of this operon, the primary structures of the four gene products have been derived and compared with the available corresponding structures from eubacteria, archaebacteria and chloroplasts. Extensive homologies were found in almost all cases and in the order 7),fus(translationelongationfactorEF−G)andtuf(translationelongationfactorEF−Tu).Fromthenucleotidesequenceofthisoperon,theprimarystructuresofthefourgeneproductshavebeenderivedandcomparedwiththeavailablecorrespondingstructuresfromeubacteria,archaebacteriaandchloroplasts.Extensivehomologieswerefoundinalmostallcasesandintheorder12 > EF-Tu > EF-G > $7; the largest homologies were generally found between the cyanobacterial proteins and the corresponding chloroplast gene products. Overall codon usage in S. platensis was found to be rather unbiased.
Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1986
The photosynthetic F1-ATPase from the cyanohacterium Spirulina platensis was recently purified in a five-suhunit, reconstitutively active form (Hicks, D.B. and Yocum, C.F. (1986) Arch. Biochem. Biophys. 245, 220-229). Here we report on the similarities of the cyanobacterial F 1 to the higher plant chloroplast F I (CF I), as judged by two distinct methods. The ability of each coupling factor to reconstitute photophosphorylation in photosynthetic membranes depleted of F I content by 2 M NaBr treatment was tested. Addition of either the homologous enzyme (e.g., Spirulina F1, Spirulina membranes) or the heterologous enzyme (e.g., spinach CF 1 Spirulina membranes) to depleted membranes increased the rate of phenazine methosulfate-dependent cyclic photophosphorylation from nearly zero to up to 70 pmol ATP/h per mg Chl. Antibodies against four subunits of CF 1 (a, 3', 8 and e) and against fl of Escherichia coli F t were reacted with the Spirulina enzyme by protein blotting. The a, fl and 3' subunits of Spirulina F l cross-reacted with antibodies against the corresponding subunits from spinach. The cross-reactivity of the ,/ suhunit correlated with previous observations that Spirulina membrane ATPase activity can he modulated by light and dithiothreitol, in a similar fashion to their effect on the enzyme from spinach chloroplasts. The ability of cyanobacterial and chloroplast enzymes to restore activity to heterologous membranes in the absence of immunological similarities between their respective 8 and e subunits may suggest that structural features other than particular amino acid sequences of these subunits are paramount in their roles in binding F~ to the membrane and in sealing proton leaks.