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31.
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
32.
Two individuals with elliptocytic red cells apparently lack three minor erythrocyte membrane sialoglycoproteins. 总被引:9,自引:4,他引:5 下载免费PDF全文
D J Anstee S F Parsons K Ridgwell M J Tanner A H Merry E E Thomson P A Judson P Johnson S Bates I D Fraser 《The Biochemical journal》1984,218(2):615-619
We have studied the erythrocytes of two individuals (P. L. and K. W.) who lack the Gerbich (Ge) blood-group antigen. The erythrocytes of P. L. and K. W. were not reactive with two monoclonal antibodies (NBTS/BRIC 4 and NBTS/BRIC 10) which reacted with normal erythrocytes. The membranes of P. L. and K. W. erythrocytes appeared to lack three minor sialoglycoproteins (beta, beta 1 and gamma). These three minor sialoglycoproteins were found to be associated with the cytoskeletons of normal erythrocytes. Approx. 10% of the erythrocytes of P. L. and K. W. were frankly elliptocytic. We suggest that one or more of the minor sialoglycoproteins may play a part in maintaining the discoid shape of the human erythrocyte. 相似文献
33.
J H Bates G K Prisk T E Tanner A E McKinnon 《Journal of applied physiology (Bethesda, Md. : 1985)》1984,56(1):254-258
A bag-in-box system (BBS) whose volume is monitored by a mechanical spirometer tends to have a slow response if the volume of the box is large, and this may significantly affect its measurement of gas flow. We describe a device for creating reproducible gas flows with which the impulse response of a BBS may be conveniently determined. Two computational techniques for correcting a BBS flow measurement for the effects of the impulse response were investigated: 1) an exponential model method that assumes a second-order model of the BBS dynamics and 2) a Fourier transform-based method of deconvolution known as Wiener filtering. Both correction methods produced a significant increase in the accuracy of BBS flow estimations, with the Wiener filter giving superior results. 相似文献
34.
Dolichylphosphate-dependent Glycosyl Transfer Reactions in the Endoplasmic Reticulum of Castor Bean Endosperm 总被引:2,自引:2,他引:0 下载免费PDF全文
In the endosperm of Ricinus communis (castor bean) a number of glycosyl transferases were found to be present during germination. They catalyze the incorporation of mannose from guanosine diphosphate mannose and of N-acetylglucosamine from uridine diphosphate N-acetylglucosamine into a glycolipid fraction, which had all of the properties of dolichylphosphate and pyrophosphate sugars, respectively. The sugar moiety of dolichylphosphate mannose is transferred to a lipid-oligosaccharide, containing more than 6 hexose units. When the membranes are preincubated with nonradioactive guanosine diphosphate mannose and uridine diphosphate N-acetylglucosamine, radioactivity from dolichylphosphate [14C]mannose is also transferred to a glycopolymer. In addition, the formation of radioactive glycoproteins from guanosine diphosphate [14C]mannose has been demonstrated using a combination of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autofluorography. 相似文献
35.
Donald P. Weidemann Robert D. Tanner Gerald W. Strandberg S.E. Shumate 《Enzyme and microbial technology》1981,3(1):33-40
It has been observed that microbial cells can adsorb uranium ions from dilute aqueous solutions. Data collected from such experiments can be used to estimate correlative mass transfer coefficients. Physical observations bear out several inadequacies, however, of using an adsorption mass transfer model with a constant transfer coefficient relating the rate of transfer to the concentration gradient. By itself, the mass transfer model contains no provision to include (1) the initial transient, (2) the curvature in the later time rate curve, and (3) the non-linear curve relating initial levels of uranium concentration in solution to final residual uranium concentration for a set of batch experiments. It is found that a better match to observed data can be achieved by utilizing an intermediate state adsorption model analogous to a kinetic model based on an enzyme - substrate coupling scheme. 相似文献
36.
The molybdenum centre of native xanthine oxidase. Evidence for proton transfer from substrates to the centre and for existence of an anion-binding site. 下载免费PDF全文
The observation by Bray & Knowles [Proc. R. Soc. London Ser. A (1968) 302, 351--353] of direct transfer, during the catalytic reaction, of hydrogen atoms from substrate molecules to the enzyme xanthine oxidase was reinvestigated. The experimental phenomenon and its basic interpretation were confirmed and extended. In the reduced functional enzyme, molybdenum(V) interacts with two enzyme-bound protons, which are exchangeable with solvent protons. One of these is coupled to the metal with AHav. 1.4mT and the other with AHav. 0.3mT. The molecule also contains a site for the binding of anions, presumably as ligands of molybdenum. This is shown by effects of nitrate ions on the e.p.r. spectra. The spectra of the nitrate and 1-methylxanthine complexes of the reduced enzyme are very similar to one another, and are designated Rapid type-1 spectra. It is concluded that, in the Michaelis complex, the substrate molecule occupies the anion site, probably being bound to molybdenum via the nitrogen in its 9-position. During the turnover process, hydrogen from the substrate C-8 position, after transfer to the enzyme, appears as the proton more strongly coupled to molybdenum. This proton then exchanges with solvent deuterium with a rate constant of 27s-1, at pH 8.2 and 12 degrees C. It has been confirmed that substrate molecules occupying the anion site do not interfere with observation of the transfer and exchange processes. 相似文献
37.
Comparison of the molybdenum centres of native and desulpho xanthine oxidase. The nature of the cyanide-labile sulphur atom and the nature of the proton-accepting group. 下载免费PDF全文
The non-functional form of xanthine oxidase known as the desulpho enzyme was compared with the functional enzyme in various ways, to obtain information on the structure of the molybdenum centre and the mechanism of the catalytic reaction. The desulpho enzyme, like the functional one, possesses a site for the binding of anions, presumably as ligands of molybdenum. Evidence is presented that in the Mo(V) e.p.r. signal from the desulpho-enzyme, as in that from the functional enzyme, a weakly coupled proton, in addition to a strongly coupled proton, interacts with the metal. Measurements were carried out by e.p.r. on the rate at which the proton strongly coupled to molybdenum exchanged, on diluting enzyme samples with 2H2O. For the desulpho enzyme the exchange rate constant was 0.40s-1, at pH 8.2 and 12 degrees C, and for the functional enzyme it was 85 s-1. It is shown that the great majority of reported differences between the enzyme forms are consistent with functional enzyme containing an (Enzyme)-Mo=S grouping, replaced in the desulpho form by (Enzyme)-Mo=O. Protonation of these groups, with pK values of about 8 and 10 respectively, would give (Enzyme)-Mo-SH and (Enzyme)-Mo-OH, these being the forms observed by e.p.r. The accepting group in the functional enzyme, for the proton transferred from the substrate while molybdenum is reduced in the catalytic reaction [Gutteridge, Tanner & Bray (1978) Biochem J. 175 869-878], is thus taken to be Mo=S. 相似文献
38.
In 18 batch-fermentation experiments, baker's yeast was grown in an enriched mineral medium, containing 10% by weight glucose, at various pH and temperature levels. The pH and temperature are just two representative engineering variables which can be easily varied at negligible cost. The commercial yeast inoculum, 20% by weight or about .16% viable cells, was selected to represent industrial (nonsterile) conditions. Free L-lysine, ethanol, and cell growth were followed in time for each batch run held at a fixed pH and temperature. The maximum free lysine level reached at either 10 1/2 or 24 hr occurred at a pH of 5 and 32 degrees C. At 24 hr, the peak free lysine level, 120 mg/liter, is three times as great as the uncontrolled pH counterpart. In terms of total L-lysine (free plus protein-bound) the peak represents a 25% improvement over the uncontrolled case, based on an average 3.5% lysine level per cell weight. The greatest measured cell level, .9% by weight in the fermentation broth, or a 5 1/2-fold increase over th inoculum, was reached during the 36 degrees C and pH 3 run, while the largest measured ethanol value (3%, or 30% conversion by weight from glucose) was achieved during the 28 degrees C and pH 6 experiment. The optimal lysine run product, however, no less than 15% of the maximum cell and 30% of the maximum ethanol levels. 相似文献
39.
Biogeographic comparisons of marine algal polyphenolics: evidence against a latitudinal trend 总被引:5,自引:1,他引:4
Nancy M. Targett Loren D. Coen Anne A. Boettcher Christopher E. Tanner 《Oecologia》1992,89(4):464-470
Summary Marine allelochemicals generally are present in greater quantity and diversity in tropical than in temperate regions. Marine algal polyphenolics have been reported as an apparent exception to this biogeographic trend, with literature values for phenolic concentrations significantly higher in temperate than in tropical brown algae. In contrast, our results, the first reported for Caribbean brown algae (orders Dictyotales and Fucales), show that many species have high phenolic levels. In addition, both our study and previous studies with north temperate and tropical species demonstrate that there is marked variation in algal phenolic levels within species from different locations. We conclude that high phenolic concentrations occur in species from both temperate and tropical regions, indicating that latitude alone is not a reasonable predictor of plant phenolic concentrations. 相似文献
40.
Two water-soluble carbodiimides, differing in molecular dimensions, have been used to characterize the cytochrome c binding site of bovine heart cytochrome c oxidase. Several polypeptide components of the enzyme contain acidic residues which are modified by these reagents. Carboxyl groups present in subunit II, VII and polypeptide c, are protected from modification when cytochrome c, equimolar to oxidase, is added and they can cross-link to the substrate once activated by the carbodiimide. Comparison of the modification patterns suggest that the most reactive residues are located on subunit II and VII, the former being also more exposed. The data obtained indicate that even though subunit II plays the major role in binding cytochrome c, at least two other lower Mr polypeptides contribute to the cytochrome c binding domain. 相似文献