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1.
R Philipp J O McIntyre B H Robinson H Huth W Trommer S Fleischer 《Biochimica et biophysica acta》1984,790(3):251-258
Calcium efflux and EGTA-induced calcium release from an internal platelet membrane fraction have been studied after the oxalate-supported calcium uptake had reached steady state. Increasing external calcium concentrations stimulate the calcium efflux velocity, with an apparent half-maximal stimulation at about 5 microM outside calcium concentration and a maximal velocity of calcium efflux of 4.66 +/- 2.32 nmol X min-1 X mg-1. Moreover, the ratio of the liberated calcium on the loaded calcium seems to be independent of the increasing external calcium concentration. Increasing the calculated internal calcium concentration by varying the oxalate potassium concentration from 10 mM to 1 mM results in an increase of the liberated calcium from the membrane vesicles from 7.4% to 63%, respectively, without changing the calcium efflux velocity. Similar conclusions can be drawn from the observation of results from the calcium efflux and EGTA-induced calcium release methods. Moreover, calcium pump reversal does not seem to be responsible for the calcium efflux or calcium release. All these different points added to the previously described regulation of calcium efflux by the catalytic subunit of cAMP protein kinase suggest us that the mechanism of calcium liberation by the platelet membranes is different from the calcium uptake. 相似文献
2.
3.
The transport of carnitine by rat kidney cortex slices against a concentration gradient has been demonstrated. Similarities to other transport systems included a linear period of uptake, as well as indications of saturability of the system with increasing concentrations of substrate. The transport of carnitine was inhibited by anoxia, and carbonyl cyanide-m-chloro-phenylhydroxazone (CCC1P), an uncoupler of oxidative phosphorylation. Carnitine uptake was stimulated approximately 50% when kidney slices were treated with dibutyryl cAMP. 相似文献
4.
Dieter Ehlers Ina Sakowski Werher Mohrig 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1989,159(4):481-489
Summary The high affinity of granulocytes of guinea pig and man to glass surfaces is modified by serum. Native serum contains both an adherence-promoting activity, which is related to complement, and components which reduce the adhesiveness of granulocytes. These components are stable at 56°C for 30 min and are tightly bound to the glass surface. -Lipoproteins are candidates for this adherence reducing ability of serum. Adherence promotion by native serum is mediated by coating the glass surface with C3b/C3bi. Human granulocytes from the peripheral blood adhered to glass surfaces coated by native human or guinea pig serum with C3b/C3bi to almost the same extent as in the presence of native serum, but on guinea pig granulocytes elicited in the peritoneal cavity, a cell surface metalloproteinase degraded the C3b/C3bi, thus reducing the adhesiveness of these cells. This proteinase was inhibited by MgEDTA, DTT, and 1,10-phenanthroline, whereby the high adhesiveness of granulocytes was restored to C3b/C3bi-coated glass.Abbreviations
BA
benzamidine hydrochloride
-
BTS
Bacillus thuringiensis subtoxicus
-
DTT
dithiothreitol
-
EAC
-amino-caproic acid
-
gp
guinea pig
-
LDL
low density lipoproteins
-
SEM
scanning electron microscopy 相似文献
5.
Jessica E. Hoogendijk Gerard W. Hensels Ina Zorn Linda Valentijn Emiel A. M. Janssen Marianne de Visser David F. Barker Bram W. Ongerboer de Visser Frank Baas Pieter A. Bolhuis 《Human genetics》1991,88(2):215-218
Summary Recently, it has been shown that Charcot-Marie-Tooth disease type 1a (CMT1a) is linked with a duplication of a DNA segment that is detected by probe VAW409R3, and that is located on chromosome 17p11.2. Here, we show that this duplication also contains VAW412R3a, but not A10-41 and EW503. Accounting for the duplication in recombination analysis, we found recombinants between CMT1a and EW301 and EW502, but not with A10-41, VAW409R3, and VAW412R3. Using pulsed-field gel electrophoresis analysis, we estimated the minimal size of the duplicated region in CMT1a patients to be 1100 kb. 相似文献
6.
The liver mitochondrial acetyl-CoA acetyltransferase (acetyl-CoA:acetyl-CoA C-acetyltransferase, EC 2.3.1.9), is involved in ketone body synthesis. The enzyme can be chemically modified and inactivated by CoASH and also by CoASH-disulfides provided glutathione is present. The unmodified enzyme shows in its denatured state 7.95 +/- 0.44 sulfhydryl groups per enzyme and in its native state 3.92 +/- 0.34 sulfhydryl groups which react with Ellmann's reagent. The modified enzyme reveals in its native state also 4.07 +/- 0.25 sulfhydryl groups per enzyme, but in its denatured state 9.10 +/- 0.51 sulfhydryl groups could be detected. Approximately four sulfhydryl groups per enzyme, unmodified or modified, can be alkylated by iodoacetamide. These results prove for each subunit the existence of two sulfhydryl groups and suggest the existence of two disulfide bridges. The CoASH modification, which should proceed at one of these disulfide groups, prevents subsequent acetylation of the enzyme and is drastically reduced in the iodoacetamide-alkylated enzyme. In the demodification of the modified enzyme, the CoASH is set free as a mixed disulfide with glutathione. 相似文献
7.
The mitochondrial acetyl-CoA acetyltransferase (acetoacetyl-CoA thiolase, EC 2.3.1.9) is involved in ketone body biosynthesis. In its unmodified state, referred to as transferase B in former publications (Huth, W. (1981) Eur. J. Biochem. 120, 557-562), the enzyme is characterized by the highest specific activity of 21.65 mumol/min per mg protein (direction of acetoacetyl-CoA synthesis); several forms of the enzyme with lower specific activities result from chemical modification by an apparent covalent binding of CoASH. The chemical modification results in an inactivation of the enzyme: a 2 h incubation with 0.2 mM CoASH at pH 8.1 at 30 degrees C inactivates up to 95%. Both processes, the CoASH-binding and the resulting inactivation, can be simultaneously reversed by treatment with glutathione. The specificity of inactivation is limited to CoASH and the intact sulfhydryl group is a prerequisite for this process. The enzyme exhibits a limited number (n = 3.2) of high-affinity (Ka = 26.7 microM) specific binding sites for CoASH. The inactivation-reactivation cycle of acetyl-CoA acetyltransferase by CoASH and glutathione may involve a protein disulfide-thiol exchange and represents a mode of control in modulating the amount of active enzyme. 相似文献
8.
Scavenging of Hydrogen Peroxide in the Endosperm of Ricinus communis by Ascorbate Peroxidase 总被引:6,自引:0,他引:6
The fat-storing endosperm of Ricinus communis L. was found tocontain an ascorbate peroxidase (EC 1.11.1.11
[EC]
), which is nearlyas active as catalase (EC 1.11.1.6
[EC]
) in degradation of hydrogenperoxide (H2O2) at its physiological concentrations. This ascorbateperoxidase probably functions together with monodehydroascorbatereductase (EC 1.6.5.4
[EC]
) or dehydroascorbate reductase (EC 1.8.5.1
[EC]
)and glutathione reductase (EC 1.6.4.2
[EC]
) to remove the H2O2 producedduring the transformation of fat to carbohydrate in the glyoxysomes.The activities of these enzymes as well as the content of ascorbateand glutathione increase parallel to the activities of glyoxysomalmarker enzymes during the course of germination. Inhibitionof catalase by aminotriazole results in increases of the ascorbateperoxidase activity and of the glutathione content. All fourenzymes are predominantly localized in the cytosol of the Ricinusendosperm with low activities found in the plastids and themitochondria. The results suggest, that the ascorbate-dependentH2O2 scavenging pathway, which has been shown to be responsiblefor the reduction of photosynthetically derived H2O2 in thechloroplasts, operates also in the Ricinus endosperm. (Received June 5, 1990; Accepted July 31, 1990) 相似文献
9.
10.
The formation of native disulfide bonds during in vitro protein folding can be limiting in obtaining biologically active proteins. Thus, optimization of redox conditions can be critical in maximizing the yield of renatured, recombinant proteins. We have employed a folding model, that of the beta subunit of human chorionic gonadotropin (hCG- beta), to investigate in vitro oxidation conditions that facilitate the folding of this protein, and have compared the in vitro rates obtained with the rate of folding that has been observed in intact cells. Two steps in the folding pathway of hCG-beta were investigated: the rate-limiting events in the folding of this protein, and the assembly of hCG-beta with, hCG-alpha. The rates of these folding events were determined with and without protein disulfide isomerase (PDI) using two different types of redox reagents: cysteamine and its oxidized equivalent, cystamine, and reduced and oxidized glutathione. Rates of the rate-limiting folding events were twofold faster in cysteamine/cystamine redox buffers than in glutathione buffers in the absence of PDI. Optimal conditions for hCG-beta folding were attained in a 2 mM glutathione buffer, pH 7.4, that contained 1 mg/mL PDI and in 10muM cysteamine/cystamine, pH 8.7, without PDI. Under these conditions, the half-time of the ratelimiting folding event was 16 to 20 min and approached the rate observed in intact cells (4 to 5 min). Moreover, folding of the beta subunit under these conditions yields a functional protein, based on its ability to assemble with the alpha subunit. The rates of assembly of hCG-beta with hCG-alpha in the cysteamine/cystamine or glutathione/PDI redox buffers were comparable (t(1/2/sb> = 9 to 12 min)). These studies show that rates of folding and assembly events that involve disulfide bond formation can be optimized by a simple buffer system composed of cysteamine and cystamine. (c) 1994 John Wiley & Sons, Inc. 相似文献