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1.
A new Dol-P-Man:protein O-D-mannosyltransferase activity from Saccharomyces cerevisiae 总被引:2,自引:0,他引:2
The deletion of the protein mannosyltransferase 1 gene (PMT1)of Saccharomyces cerevisiae results in viable cells. O-Mannosylationof proteins is reduced to about half of the value in comparisonto wild-type cells. In order to distinguish between the thePMT1 gene product (= Pmt1p) and residual transferase activity,an in vitro assay to measure Dol-P-Man:protein mannosyltransferaseactivity in cells deleted for PMT1 has been developed. The transferaseactivity of these cells exhibits a pH optimum of 6.5 as comparedto pH 7.5 for Pmt1p. The K$$$ value of the residual enzyme activityfor the hexapeptide YNPTSV is 7 times higher than that of Pmt1pand shows a clear preference for the seryl/residue. Differencesin substrate affinities as well as in seryl/threonyl preferencebetween the two enzymes, however, depend on the specific sequenceof the peptides used in the enzyme assay. The new enzyme activityshows a significantly lower thermal stability as compared toPmt1p. glycoprotein O-glycosylation mannosyltranferase Saccharomyces cerevisiae 相似文献
2.
3.
Chalcone synthase (CHS), a key enzyme of flavonoid synthesis,was localized in shoot apices of Fagopyrum, Brassica and Pisum.The enzyme was detected in initial cells of the shoot apex,which gives rise to the whole plant body. In Fagopyrum and BrassicaCHS was located in the rib and flank meristems, whereas in theArgenteum mutant of Pisum this enzyme was localized at an earlierstage in the ontogenesis of the shoot. It occurs in a few cellsof the tunica, which gives rise to the protoderm, and then tothe epidermis which contains anthocyanins in these plants. Chalcone synthase, immunogold labelling, promeristem, shoot apex, Brassica, Fagopyrum, Pisum 相似文献
4.
In synchronized Chlorella sorokiniana cells, the NH4+ inducibleNADP-specific glutamate dehydrogenase enzyme (NADP-GDH) accumulatedin a linear manner throughout the first cell cycle. Early inthe following second cell cycle, an increase in its rate ofaccumulation occurred that was proportional to the increasein total cellular DNA in the previous cell cycle. In synchronizedbacterial cells, increases in rate of linear accumulation ofinducible enzymes coincide with the time of replication of theirstructural genes. To determine whether the rate change in NADPGDHaccumulation resulted from a delay in replication of its nuclearstructural gene (gdhN) in fully induced C. sorokiniana cells,the cell cycle timing of replication of this gene was comparedto that of another nuclear gene, nitrate reductase (nia), andof a chloroplast gene, ribulose bisphosphate carboxylase large-subunit(rbcL), in synchronized cells cultured in NH4+ or NO3(uninduced) medium. The gdhN and nia genes replicated withinthe period of nDNA synthesis and rbcL within the period of ctDNAsynthesis in cells growing in either nitrogen source. Therefore,the delayed rate change in enzyme accumulation results froma process that regulates expression of the gdhN gene after itsreplication. (Received July 16, 1994; Accepted November 28, 1994) 相似文献
5.
The Dichotomiflora group of Panicum contains NAD-malic enzyme(ME) species with centrifugal chloroplasts in Kranz cells, NAD-ME(F)species as well as NAD-ME species with centripetal chloroplastsin Kranz cells, NAD-ME (P) species. Many attributes of leafanatomy of 22 C4 Panicum species were investigated to identifydifferences among four different C4 subtypes, i.e. NADP-ME,NAD-ME(F), NAD-ME(P) and PEP-CK species grouped by the C4-aciddecarboxylating enzymes and chloroplast location in Kranz cellsin combination. Differences were found in the number of Kranzcells surrounding a large vein, and the number surrounding asmall vein, the interveinal distances, the proportion of leafcross sectional area occupied by epidermis plus sclerenchyma,by mesophyll cells, by Kranz cells, and by vascular bundles.There were also differences in the ratios of the area of thedifferent cell types. The number of the characters significantlydifferent between a respective pair of C4 subtypes was the largestbetween NAD-ME(F) and NAD-ME(P) species. In principal componentanalysis applied to 11 leaf anatomical characters, the differentC4 subtypes clustered into small groups, although the rangeof variations of PEP-CK species and those of NAD-ME(F) speciesoverlapped. The results were discussed in relation to taxonomyand ecological adaptation of Panicum species in the differentC4 subtypes. C4 photosynthesis, NADP-malic enzyme, NAD-malic enzyme, Phosphoenolpyruvate carboxykinase, C4 leaf anatomy, Panicum, Kranz, Dichotomiflora group 相似文献
6.
The specificities for cytochrome c of the aa3-type cytochromec oxidase were studied with enzymes derived from Thiobacillusnovellas, Nitrobacter agilis, Paracoccus denitrificans and thecow in reaction with the cytochromes c from 5 prokaryotes and7 eukaryotes. The T. novellus enzyme reacted most rapidly withthe cytochromes c of Candida krusei, tuna and bonito as wellas T. novellus cytochrome c; the specificity for cytochromec of the N. agilis enzyme was similar to that of the T. novellusenzyme. The bovine enzyme reacted rapidly with all the eukaryoticcytochromes c tested. The P. denitrificans enzyme showed a specificitysimilar to that of the bovine enzyme, except that it reactedrapidly with P. denitrificans cytochrome c, while the bovineenzyme reacted with it very poorly. All four kinds of enzymesshowed an extremely limited reaction with Pseudomonas aeruginosacytochrome c. The amino acid composition of subunit I of the N. agilis enzymeresembled that of the bovine enzyme, while the compositionsof their subunits II were different. On the basis of these results,an evolutionary relationship between bacterial and eukaryoticenzymes was discussed. (Received May 21, 1981; Accepted August 20, 1981) 相似文献
7.
The greatest activity of protopectinase obtained from the growthof Rhizoctonia solani and Fusarium solani on autoclaved potatoplugs occurred at pH 6.5, and greatest activity of the lossof viscosity enzyme was found at 65 for Rhizoctonia,and between 6.5 and 8.3 for Fusarium. Protopectinase enzymeobtained from double infections of the Fusarium spp. with Rhizoctonia,or by mixing the enzymes of individual Fusarium spp. with Rhizoctoniaenzyme, were more active than the enzymes from single inoculations.Cylindrocarpon radicicola enzyme was more active when obtainedfrom a pure culture than from double infection. Similarly, mixingthis enzyme with the enzyme of Rhizoctonia reduced its activity.The evidence indicated that the protopectinase of Rhizoctoniawas similar to that of Cylindrocarpon and differed from thatof the Fusarium spp. Using paper partition chromatography, two bands from Rhizoctoniacrude enzyme had a stimulatory effect on Fusarium enzyme, whileonly one band from Fusarium enzyme stimulated Rhizoctonia enzyme. The purified enzyme of Rhizoctonia degraded pectin to galacturonicacid. Fusarium pure enzyme degraded pectin to an intermediatestage. A mixture of the two enzymes degraded pectin to galacturonicacid, without the intermediate stage formed by Fusarium alonebeing detected. The role played by pectic enzymes upon the synergistic relationof Rhizoctonia solani and Fusarium solani on rotting potatotubers is discussed. 相似文献
8.
1. The hydrogenase reactions in a purple non-sulfur bacterium,Rhodopseudornonas palustris, were investigated. Under photoheterotrophicculturing conditions, the photosynthetic activity of the cellswas found to be closely paralleled by the activity of hydrogenase.It was also revealed that the bacterium can grow under suchconditions even withont both photosynthetic and the enzyme activities. 2. The enzyme was revealed to be localized in the particulatefraction (presumably chromatophores) of the disintegrated cells.The properties of the enzyme in the cell-free preparation andin intact cells were described. 3. Among various hydrogen acceptors tested, p-benzoquinone wasmost rapidly hydrogenated. Some heat-labile factor was shownto be involved in the reduction of quinone, which was not requiredin the reduction of methylene blue. 4. The reactions of hydrogenase, both in cell-free state (quinone-and MB-reduction) and in intact cells (oxyhydrogen reaction),were markedly inhibited by molecular oxygen. The inhibitionwas noncompetitive with respect to H2. A reversible mole-to-molecombination of the enzyme and O2 was suggested as the mechanismof the inhibition. 5. Carbon monoxide inhibition was suggested also to be causedby a reversible mole-to-mole combination of the enzyme and theinhibitor. This inhibition was competitive with respect to H2. 6. Rhodopseudomonas palustris hydrogenase was found to be refractorytoward cyanide, azide and sulfhydryl reagents. 7. Light markedly suppressed the oxyhydrogen reaction (intactcells) whereas other hydrogenation reactions (intact cells andcell- free preparations) were not affected by illumination.
1Present address: Laboratory of Biological Chemistry, TokyoInstitute of Technology, Meguro-ku, Tokyo. (Received August 21, 1961; ) 相似文献
9.
Chlorophyllase was extracted from green cells of Chlorella protothecoidesby n-butanol treatment and purified 600-fold, as measured byenzyme activity in chlorophyll a hydrolysis, by ammonium sulfateprecipitation, chromatography on TEAE-cellulose column and gelfiltration with Sephadex G-200. At each purification step the following activities were compared:hydrolyses of chlorophyll a and methyl chlorophyllide a, methanolysisof chlorophyll a and transphythylation of methyl chlorophyllidea to chlorophyll a. The ratio of activities of chlorophyll a hydrolysis to chlorophylla methanolysis changed on purification and partial inactivationby heat, PCMB and phytol, as well as by varying the reactiontemperature, thus suggesting that the two reactions are notcatalyzed by a single enzyme. In contrast, the activity ratio of chlorophyll a methanolysisto transphytylation of methyl chlorophyllide a remained unaltered,indicating that these reactions can be forward and backwardreactions catalyzed by one enzyme. Results of kinetic studies also indicated that the chlorophyllaseof Chlorella protothecoides consists of at least two enzymes.One enzyme catalyzes chlorophyll a hydrolysis and the other,chlorophyll a methanolysis and the reverse reaction, transphytylationof methyl chlorophyllide a. (Received May 24, 1973; ) 相似文献
10.
Isoamyl acetate, one of the major characteristic aroma compoundsof banana fruit (Musa sapientum L.), was synthesized by thecondensation of acetyl-CoA and isoamyl alcohol under the actionof alcohol acetyltransferase, which was found to be localizedin the soluble fraction of pulp cells. The activity of thisenzyme increased with the ripening of banana fruit. The enzyme was purified about 62-fold. The purified enzyme wasvery labile at pHs lower than pH 7.0 but relatively stable atpH 7.5{small tilde}9. The enzyme was most active at 30C andpH 8.5. The molecular weight was estimated to be about 40,000by gel filtration. Its Km values for acetyl-CoA and isoamylalcohol were 50 µM and 0.4 mM, respectively. (Received January 28, 1985; Accepted May 30, 1985) 相似文献
11.
The green alga Euglena gracilis contains a thiol methyltransferasethat catalyzes the S-adenosylmethionine-dependent methylationof pentachlorobenzenethiol. The enzyme was localized in thecytoplasm and partially purified. The pH optimum for the enzymewas 6.5. The enzyme methylated a number of foreign thiols, butnot the cellular thiols, glutathione or cysteine. Phenols andanilines were not substrates. When pentachloro-benzenethiolwas the methyl acceptor the Km was found to be 82 µM andthe corresponding Km for S-adenosylmethionine was 140 µM.The molecular weight of the enzyme was 21,000, as determinedby gel filtration. A role for this enzyme in detoxifying xenobioticthiols is proposed. (Received September 28, 1984; Accepted April 25, 1985) 相似文献
12.
1. From nitrate-adapted cells of Rhodospirillum rubrum, an activepreparation of nitrate reducing enzyme was isolated in partiallypurified state. The enzyme was found to be localized in thechromatophores of the cell and, on sonication, readily releasedinto the upernatant fraction. The purified enzyme, catalyzingthe electron transfer between DPNH and nitrate, contained ab-type cytochrome, flavin and non-heme iron, which was removedon dialysis in the presence of cyanide. Besides DPNH, only methylviologen(reduced form) was effective as electron donor. 2. The effects of pH and the addition of various activatorsand inhibitors on the rate of nitrate reduction were investigated,using DPNH or reduced methylviologen as the electron donor.The oxidation-reduction of the flavin and the heme in the enzymewas followed spectrophotometrically. A pathway of electron inthe nitrate reduction through this enzyme was proposed. 3. The nitrate reductase of this bacterium was compared withother nitrate reductases obtained from other sources, and themetabolic roles of this enzyme were discussed. In the nitrate-adaptedcells of Rsp. rubrum, only one and the same enzyme was obtainedunder different growth conditions of nitrate assimilation (i.e., nitrate as N-source; light as energy source) and nitrate-respiration(i. e., in the dark; nitrate as hydrogen acceptor and N-source).
1 Dedicated to Prof. H. TAMIYA on the occasion of his 60th birthday.This paper was submitted to the University of Tokyo to fulfillthe requirement for the author's doctorate.
2 Present address; Botanical Institute, Kyoto University. (Received December 14, 1962; ) 相似文献
13.
Ultrastructural and functional differences between the crossveins of Digitaria eriantha and Zea mays were investigated.Cross veins of both genera possess similar conducting tissues,namely one tracheary element and one sieve cell. In Digitariaeriantha these conducting elements are associated with onlytwo parenchyma cells, and, those in Zea mays are completelysurrounded by chJorenchyma cells. The protein ribulose 1,5-bisphosphatecarboxylase/oxygenase (rubisco) was used as a probe for CO2fixation sites by comparing its distribution in the varioustissue types in the leaves of the two genera. The protein wasfound to be equally and uniformly distributed in the stromalregions of the chlorenchyma sheath cell chloroplasts of longitudinalveins of both genera. The chlorenchyma sheath cells in crossveins of Zea mays show a similar distribution of the enzymeas the longitudinal bundles. However, this enzyme was shownto be absent in the cross vein parenchyma cells of Digitariaeriantha and in the mesophyll cells of both genera. Cross veins, immuno-gold labelling, ribulose 1,5-bisphosphate carboxylase/oxygenase, Digitaria eriantha, Zea mays 相似文献
14.
Cells of Porphyridium cruentum R-l, a unicellular red alga,grown under ordinary air (0.04% CO2) showed much higher activityof carbonic anhydrase (CA) than those grown under CCvenrichedair (2% CO2). CA activity was not detected in a suspension ofintact cells, and was detectable only after the cells had beenhomogenized, indicating that this enzyme was localized onlywithin the algal cells. After partial purification of Porphyridium CA, its mol wt wasestimated as 59 kDa by SDS-PAGE and 55 kDa by gelfiltration.This suggests that the native enzyme is a monomer. Its activitywas not affected by benzensulfonamides, potent inhibitors ofCAs isolated from Chlamydomonas and other organisms. Chloride(or bromide) ions was essential for CA activity. CA activitymarkedly decreased when the cell extract had been incubatedat pH lower than 7 before assay. Upon readjusting the pH ofthe preincubation medium to 9 or higher, the enzyme activitywas restored, indicating that the inactivation is reversible. (Received April 17, 1987; Accepted July 21, 1987) 相似文献
15.
The Involvement of Glucose-6-Phosphatase in Mucilage Secretion by Root Cap Cells of Zea mays 总被引:1,自引:0,他引:1
In order to determine the involvement of glucose-6-phosphatasein mucilage secretion by root cap cells, we have cytochemicallylocalized the enzyme in columella and peripheral cells of rootcaps of Zea mays. Glucose-6-phosphatase is associated with theplasmalemma and cell wall of columella cells. As columella cellsdifferentiate into peripheral cells and begin to produce andsecrete mucilage, glucose-6-phosphatase staining intensifiesand becomes associated with the mucilage and, to a lesser extent,the cell wall. Cells being sloughed from the cap are characterizedby glucose-6-phosphatase staining being associated with thevacuole and plasmalemma. These changes in enzyme localizationduring cellular differentiation in root caps suggest that glucose-6-phosphataseis involved in the production and/or secretion of mucilage byperipheral cells of Z. mays. Zea mays, corn, glucose-6-phosphatase, columella cell, peripheral cell, mucilage, secretion, cytochemistry 相似文献
16.
Commercially available cell wall-degrading enzymes frequentlyused for protoplast isolation inhibited CO2 fixation and photosyntheticO2 evolution, and stimulated dark respiration by leaf tissueand isolated mesophyll protoplasts of Nicotiana tabacum L. andAntirrhinum majus L. They also depolarized the membrane potentialof cells of leaf tissue, inhibited uptake of 86Rb by tobaccoleaf tissue and isolated mesophyll protoplasts, and stimulated36CI uptake by tobacco leaf tissue. Where studied, these effectswere found to be reversible. The depolarization effect on Antirrhinumleaf cells occurred even when the enzyme preparations had beendenatured, dialysed, or desalted, and the effect was greatestin those fractions of the enzyme preparation which showed thehighest cellulase activity. Plasmolysis of tobacco leaf tissue inhibited photosyntheticO2 evolution, CO2 fixation, and 86Rb uptake to levels belowthose exhibited by isolated protoplasts in media of the samecomposition and osmolarity. The implications of these resultsfor work with leaf tissue and isolated protoplasts are discussed. 相似文献
17.
Illuminating a colorless mutant of Chlorella vulgaris 11h (M125)with blue light caused a reversible photoreduction of b-typecytochrome, i.e., absorbance increases at 423, 525 and 557 nm.This light-induced reduction of cytochrome b was most pronouncedin nitrate-starved cells, which showed some blue light responsesin carbon metabolism, including enhancement of respiration byblue light as reported previously. Prolonged illumination withblue light caused a decrease in the rate of the reduction. The photoactivation of nitrate reductase in the mutant cellswas studied in both cell-free crude extract and purified enzyme.The absorption spectrum of purified enzyme showed three peaksat 423, 525 and 557 nm after the addition of a reductant, indicatingthat the spectrum is that of cytochrome b associated with nitratereductase. Nitrate reductase activity was easily enhanced byblue light illumination after 1 min; red light had no effecton it. The blue light activation of nitrate reductase was notsignificant in growing cells, which showed its high activity. The relationship between the blue light-induced reduction ofcytochrome b and carbon metabolism is discussed. (Received September 30, 1987; Accepted February 9, 1988) 相似文献
18.
Daily analysis of anther samples during flower development hasenabled an estimation of the duration of defined developmentalperiods in pollen of the grass Phalaris tuberosa. A similarsequence of pollen development has been established for ryegrass,Lolium perenne, where changes in activity of wall enzymes havebeen followed using quantitative cytochemical methods. Acidphosphatase, an intine enzyme, showed two periods of activity:during the vacuolate period corresponding to deposition of theintine polysaccharides; and in the maturation period correspondingto cytoplasmic activity. Non-specific esterase showed greatestactivity in the parietal tapetal cells until their dissolutionearly in the vacuolate period when an increase in pollen-associatedactivity occurred. These changes provide additional evidencefor the transfer of tapetal proteins to exine sites. Lolium perenne L., Phalaris tuberosa L., ryegrass, canary grass, pollen development, quantitative cytochemistry, enzyme activities, acid phosphatase, esterase 相似文献
19.
Tanaka Katsuyuki; Takio Susumu; Yamamoto Isamu; Satoh Toshio 《Plant & cell physiology》1996,37(4):523-529
Suspension-cultured cells of Marchantia paleacea var. dipteracontain a single form of CuZn-superoxide dismutase (SOD; EC1.15.1.1
[EC]
) which is localized in the cytosol. SOD activity wasfound in cells cultured under heterotrophic, photoheterotrophicand photoautotrophic conditions. The CuZn-SOD was purified tohomogeneity from liverwort cells that had been cultued hetertrophically.Its molecular mass was 32.6 kDa, and it contained 17.5 kDa subunits,an indication that the enzyme is a homodimer. The enzyme hadpeaks of absorption at 252, 258 and 264 nm in the ultravioledregion, due to the presence of phenylalanine, and a peak at680 nm in the visible region, which is characteristic of CuZn-SODsfrom cholorplasts. The amino acid sequence of the amino-terminalregion of the enzyme exhibited a very high degree of homologyto those of cholorplast CuZn-SODs. An antiserum raised againstthe CuZn-SOD from liverwort cross-reacted more strongly withthe enzyme from spinach chloroplasts, than with the enzyme fromspinach cytosol. These results indicate that the CuZn-SOD ofliverwort resembles CuZn-SOD in chloroplasts even though theformer is located in the cytosol. (Received November 27, 1995; Accepted April 5, 1996) 相似文献
20.
The sheathing leaf bases of Syrrhopodon texanus are primarilycomposed of porose, thin-walled cells called hyalocysts. Largepores develop in several of the hyalocyst walls through thegradual removal of wall material. A cellulase with a pH optimumof 4.5 was detected in extracts of Syrrhopodon gametophoresby viscometric assays and dinitrosalicylic acid assay for reducingsugars. Cytochemical localization of cellulase activity in associationwith thinning hyalocyst cell walls implicate this enzyme inpore formation and is the first direct evidence of cellulaseactivity in a member of the Bryophyta. Syrrhopodon texanus, cellulase, cytochemistry, Bryophyta, hyalocyst 相似文献