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1.
2.
Location of a dicyclohexylcarbodiimide-reactive glutamate residue in the Neurospora crassa plasma membrane H+-ATPase 总被引:3,自引:0,他引:3
M R Sussman J E Strickler K M Hager C W Slayman 《The Journal of biological chemistry》1987,262(10):4569-4573
The proton pump (H+-ATPase) found in the plasma membrane of the fungus Neurospora crassa is inactivated by dicyclohexylcarbodiimide (DCCD). Kinetic and labeling experiments have suggested that inactivation at 0 degrees C results from the covalent attachment of DCCD to a single site in the Mr = 100,000 catalytic subunit (Sussman, M. R., and Slayman, C. W. (1983) J. Biol. Chem. 258, 1839-1843). In the present study, when [14C]DCCD-labeled enzyme was treated with the cleavage reagent, N-bromosuccinimide, a single major radioactive peptide fragment migrating at about Mr = 5,300 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was produced. The fragment was coupled to glass beads and partially sequenced by automated solid-phase Edman degradation at the amino terminus and at an internal tryptic cleavage site. By comparison to the DNA-derived amino acid sequence for the entire Mr = 100,000 polypeptide (Hager, K., and Slayman, C. W. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 7693-7697), the fragment has been identified as arising by cleavage at tyrosine 100 and tryptophan 141. Covalently incorporated [14C]DCCD was released at a position corresponding to glutamate 129. The DCCD-reactive glutamate is located in the middle of the first of eight predicted transmembrane sequences. When the sequence surrounding the DCCD site is compared to that surrounding the DCCD-reactive residue of two other proton pumps, the F0F1-ATPase and cytochrome c oxidase, no homology is apparent apart from an abundance of hydrophobic amino acids. 相似文献
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4.
Christiane Wloczyk Achim Kröger Thomas Göbel Gabriele Holdt Ralf Steudel 《Archives of microbiology》1989,152(6):600-605
Wolinella succinogenes grown on formate and elemental sulphur was found to use the polysulphide derivatives 2,2-tetrathiobispropionate (R2S4) or pentathionate (S5O
6
=
) as acceptors for formate oxidation. The specific activities of formate oxidation with these acceptors were similar to those with elemental sulphur. The main reaction products of R2S4 reduction were 2,2-dithiobispropionate (R2S2) and sulphide. Pentathionate was converted to thiosulphate and some elemental sulphur. The electrochemical proton potential
across the cytoplasmic membrane of the bacterium was measured in the steady state of electron transport from formate to R2S4. The electrical proportion () of the
determined through the distribution of labeled tetraphenylphosphonium cation was obtained as 0.17 Volt. The was zero, when a protonophore was present. The pH-difference across the membrane was negligible. Thus the
generated by sulphur respiration is close to that measured earlier with fumarate as the terminal acceptor of electron transport.Abbreviations DMO
5,5-dimethyloxazolidine-2,4-dione
- R2Sn (n=2–5)
2,2-polythiobispropionate
- TTFB
4,5,6,7-tetrachloro-2-trifluoromethylbenzimidazol
- TPP
tetraphenylphosphonium cation 相似文献
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6.
Zusammenfassung Anhand einer 16jährigen Untersuchung im Raum Ostwestfalen/Bielefeld wird der Bruterfolg des Turmfalken anhand von 439 Gelegen und 2256 Eiern beschrieben. Drei Brutplatztypen können unterschieden werden: A. Baumbruten in Nestern; B1. Baumbruten in Nistkästen; B2. Gebäudebruten in Nischen oder Nistkästen. Zwischen Baumbruten (A) und Nistkastenbruten (B1/2) werden signifikante Unterschiede beschrieben, die für Nistkästen größere Gelege (ca. ein Ei mehr) und größeren Ausfliegeerfolg belegen. Zwischen Nistkästen in Bäumen (B1) oder an Gebäuden (B2) konnten keine signifikanten Unterschiede festgestellt werden. Weiterhin werden Lege- und Schlupftermine, Legerhythmus und oologische Daten aus dem Untersuchungsgebiet angegeben.
Breeding biology of Kestrels (Falco tinnunculus) in Eastern Westfalia 1972–1987
Summary The 16 years of study gave 439 clutches with 2,256 eggs. We separated three types of breeding sites: the use of (a) stick nests, mostly built by corvids (cf. Tab., Fig. 3), (b1) nest boxes attached to trees or telegraph poles (Fig. 2) and (b2) nest boxes or cavities at or in buildings (Fig. 1). Within these different types of breeding places we found some significant differences. Stick nests had less eggs and though less breeding success, which was possibly caused by predation of corvids, especially magpies. Within the two types of places with nest boxes no significant differences could be established. We concluded, that stick nests were marginal in Kestrels and nest boxes were optimal despite of their placement in trees, at poles or in buildings. Furthermore, the timing of breeding cycle was described (Fig. 4) and laying interval was determind to an average value of approximately two days (Fig. 5). Mean egg size was and average volume 21.2 cm2. Two daily controlled clutches lost 15.5% and 16.1% of mass (Fig. 6) pressumably mostly due to water losses.相似文献
7.
8.
H+-translocating ATPase and pyrophosphatase (PPase) associatedwith the tonoplast of Chara corallina were isolated with theaid of a perfusion technique, and the effects of ions on theiractivities were studied. All the alkali metal cations testedstimulated the ATPase and ATPdependent H+ pumping activitiesonly by 10 to 40%. Anions, on the other hand, strongly affectedthe activities. Potassium salts of Cl- and Br- stimulated them,while F- and NO3- inhibited them. By contrast, the H+-translocatingPPase was insensitive to anions but sensitive to cations. Theorder of cation stimulation was Rb+=K+>Cs+>Na+=Li+>choline+.NO3- (50 mil), thought to be a specific inhibitor of the tonoplast-typeH+-ATPase, inhibited the ATPdependent H+ pumping almost completelybut the ATPase activity by only about 50%. Na+ inhibited thePP1-dependent H+ pumping (I5O=5OmM) in the presence of 50 mMKCl but not the ATP-dependent one. The PPase was more sensitiveto F- (I50=400µM) than the ATPase. Both the H+-ATPaseand the H+-PPase required Mg2+ for their activities, althoughan excess was inhibitory to both. The different sensitivitiesof the PP1-dependent and the ATP-dependent H+- pumping enzymesto ions correspond to the tonoplast enzymes of higher plantsand may be used as "markers" to distinguish between these enzymesin characean cells (Received October 2, 1987; Accepted May 18, 1988) 相似文献
9.
Characterization of the H Translocating Adenosine Triphosphatase and Pyrophosphatase of Vacuolar Membranes Isolated by Means of a Perfusion Technique from Chara corallina 总被引:2,自引:2,他引:0
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Sealed tonoplast vesicles were isolated from single cells of Chara corallina with the aid of an intracellular perfusion technique in combination with a 3/10% Percoll two step gradient centrifugation. The isolated tonoplast fraction was free from plasmalemma and chloroplasts, and showed no activities of cytochrome c oxidase, and latent IDPase, but had about 10% of the NADH-cytochrome c reductase activity. The vesicles had both ATPase and PPase activities, which could be stimulated in the presence of 10 micromolar gramicidin by 170 and 130%, respectively, demonstrating the existence of sealed vesicles. Furthermore, ATP- and PPi-dependent H+ pumping through the membrane into the vesicles was shown. Both ATPase and PPase had pH optima around pH 8.5. At the physiological pH, 7.3, they still had more than 80% of their maximal activities. Ammonium molybdate, azide, and vanadate had no or little effect on the activities of both enzymes or their associated H+ pumping activities. N,N′-dicyclohexylcarbodiimide inhibited the ATPase strongly (I50 = 20 micromolar) but the PPase only weakly. The ATPase was also more sensitive to N-ethylmaleimide than the PPase. 4,4′-Stilbenedisulfonic acid affected both enzyme activities and their associated H+ pumping activities. This is in contrast to the H+-PPase of higher plants which is 4,4′-stilbenedisulfonic acid insensitive. 相似文献
10.
Identification and characterization of an {alpha}-mannosidase from Trypanosoma cruzi 总被引:1,自引:0,他引:1
Swanson Paul M.; Carter Clint E.; Hager Cindy; Kim Wan Joon; Obermeier Sarah; Oeltmann Thomas N. 《Glycobiology》1992,2(6):563-569
In this report we describe the first purification and characterizationof the acid -mannosidase from the human parasite Trypanosomacruzi. The purified enzyme exhibited a native mol. wt of 240000 Da and is apparently composed of four identical subunitsof mol. wt 58 000 Da. Each of the four subunits contains oneN-linked high-mannose-type oligosaccharide. The -mannosidaseexhibited a pH optimum of 3.5 and a pI of 5.9. This low pH optimumand the ability of swainsonine to inhibit its activity suggestthat the -mannosidase is a lysosomal enzyme. Antibodies againstthe T.cruzi enzyme did not react with mammalian lysosomal -mannosidaseand, conversely, antibody against a rat lysosomal -mannosidasedid not react with the T.cruzi enzyme. Thus, the T.cruzi enzymeappears to be distinct from its mammalian counterpart. -mannosidase lysosomal enzyme Trypanosoma cruzi 相似文献