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71.
Uri S. Ladror Gary T. Wang William L. Klein Thomas F. Holzman Grant A. Krafft 《Journal of Protein Chemistry》1994,13(4):357-366
Fluorogenic peptide substrates designed to encompass the reported-secretory and amyloidogenic cleavage sites of the amyloid- precursor protein (PP) were used to analyze proteinase activities in brain extracts from control patients and those with Alzheimer's disease (AD). Activity against the secretory substrate atpH 7.5 in control and AD brains produced a major endopeptidase cleavage at the Lys687-Leu688 bond (PP770 numbering), consistent with thePP secretase cleavage. Activity in control brains against the amyloidogenic substrate atpH 7.5 produced one cleavage at the Ala673-Glu674 bond, two residues C-terminal to the amyloidogenic Met-Asp site. However, in three of four AD brains, the major cleavage was at the Asp-Ala bond, one residue from the amyloidogenic site. Both endopeptidase and carboxypeptidase activities in AD brains were lower than in control brains. Proteinase activities against the secretory substrate had a major optimum atpH 3.0–4.0 and another atpH 6.0–7.5. Proteinase activities against the amyloidogenic substrate had a major optimum at or belowpH 3.0 and another atpH 6.0. Using both substrates, activities at lowpH were higher in AD brains than in controls, while atpH above 6.5, activities in control brains were higher than in AD. These results indicate that the levels of proteolytic enzymes in AD brains are altered relative to controls.Abbreviations A
Amyloid-
- ACN
acetonitrile
- AD
Alzheimer's disease
- PP
amyloid- precursor protein
- DABCYL
4-(4-dimethylaminophenylazo)-benzoic acid
- EDANS
5-{(2-aminoethyl)amino}napthalene-1-sulfonic acid
- MES
morpholinoethane sulfonic acid
- MOPS
morpholino-propane sulfonic acid
- RP-HPLC
reverse-phase high-performance liquid chromatography
- SDS-PAGE
sodium do-decyl sulfate-polyacrylamide gel electrophoresis
- TFA
tri-fluoroacetic acid
- Tris
tris(hydroxyethyl)aminomethane 相似文献
72.
We have previously shown that micromolar concentrations of GDP stimulate the GTP-mediated phosphorylation of p36, the subunit of succinyl-CoA synthetase (SCS), in lysates prepared fromDictyostelium discoideum. In this study, we report that this phenomenon represents an enhanced catalytic capacity of SCS to form the phosphoenzyme intermediate. Low concentrations of GDP stimulate phosphoenzyme formation by either GTP, or succinyl-CoA and Pi. Under these conditions GDP enhances the apparent rate of phosphoenzyme formation but does not significantly alter the fraction of phosphorylated enzyme. This effect is retained during purification of the protein and is also observed with purified pig heart SCS, indicating that GDP directly alters the enzyme to enhance its rate of phosphorylation. Under these conditions, GDP does not function at the catalytic site, implying an allosteric regulation of SCS.Abbreviations used SCS
succinyl-CoA synthetase
-
P
i
inorganic phosphate
- NDP
nucleotide diphosphate
- NTP
nucleotide triphosphate
- PFK
phosphofructokinase A-form; ADP-forming SCS; G-form; GDP-forming SCS 相似文献
73.
We used a computer-assisted morphometry approach to analyze quantitatively the process of cytoplasmic granule formation in mouse pancreatic acinar cells stimulated with pilocarpine to induce secretion. Our findings suggest that each condensing vacuole/immature granule of pancreatic acinar cells is formed by the progressive aggregation of 106 to 128 unit progranules of narrowly fixed volume, define a range of 7.7 to 9.2 for the factor of volume condensation between the largest immature granules and the mature unit granule, and predict that the formation of a single mature unit granule by the aggregation and fusion of unit progranules involves a net reduction of at least 95% in the amount of membrane surface area associated with these structures. 相似文献
74.
Many deterministic models of sexually transmitted diseases, as well as population models in general, contain elements of stochastic or statistical reasoning. An example of such a model is that of Dietz and Hadeler (1988) concerning sexually transmitted diseases in which there is partnership formation and dissolution. Among the interesting formulas in this paper, which enter into the analysis of the model, are those for the expected number of partners a male or female has during a lifetime. To a probabilist such formulas suggest the possibility that some stochastic process may be constructed so as to yield these formulas as well as others that may be of interest. The principal purpose of this paper is to demonstrate that such a stochastic process does indeed exist in the form of a three state semi-Markov process in continuous time with stationary laws of evolution and with a one-step density matrix determined by four parameters which were interpreted as constant latent risk functions in the classical theory of competing risks. This construction of a semi-Markov process not only provides a framework for the systematic derivation of the formulas of Dietz and Hadeler but also suggests pathways,for extensions to the age-dependent case.This research was partially supported by NATO Grant D.890350 相似文献
75.
Leucrose formation from sucrose and fructose by dextransucrase is of practical interest. It has been investigated at different experimental conditions, including the influence of temperature on reaction rate and selectivity. Under appropriate conditions high product yield can be obtained. Furthermore, a model is presented that allows interpretation of the experimental data. 相似文献
76.
It has been proposed that peg formation in the vascular transition region (TR zone) between the hypocotyl and the root in
Cucurbitaceae seedlings is a gravimorphogenetic phenomenon. Initiation of the peg became visible 36 h after imbibition when
cucumber (Cucumis sativus L. cv. Burpee Hybrid II) seeds were germinated in a horizontal position at 24°C in the dark. Simultaneously, sedimented amyloplasts
(putative statoliths) were apparent in the sheath cells surrounding the vascular strands, and in the cortical cells immediately
adjacent to them, in the TR zone. In contrast, the other cortical cells, some of which were destined to develop into the peg,
contained amyloplasts which were not sedimented. These results suggest that the graviperception mechanism for peg formation
may be like that of statoliths in shoot gravitropism. By 48 h following imbibition, the cells of the TR zone still had sedimented
amyloplasts but had lost their sensitivity to gravity, possibly because of their maturation. 相似文献
77.
Tetsuko Noguchi 《Protoplasma》1994,180(1-2):29-38
Summary The formation and the decomposition of vacuoles in a member of Xanthophyceae,Botryococcus braunii, were examined by light and electron microscopy. Particles around the nucleus were identified as vacuoles from their stainability with neutral red. These particles disappeared during cell division. They reacted positively in an activity test for acid phosphatase. Electron microscopy revealed the presence of spherical vacuoles around the nucleus. During cell division, these vacuoles seemed to be decomposed by the ER which surrounded the vacuoles. Soon after this decomposition, many immature multivesicular bodies (MVBs) appeared to develop from the trans-Golgi network (TGN) and were pinched off from the TGN. These immature MVBs took up small vesicles in them as they grew into the mature MVBs. Mature MVBs took up and digested the surrounding cytoplasm, fused with one another, and eventually became the vacuoles.Abbreviations MVB
multivesicular body
- TGN
trans-Golgi network 相似文献
78.
Selenite has been shown to undergo intracellular metabolism that results in its conversion to other low molecular weight Secontaining
species and also to its incorporation into a selenocysteine residue in selenoprotein. In order to investigate whether the
incorporation into protein is required for the cytotoxic effects of selenite, we have examined whether inhibition of protein
synthesis prevents the inhibitory effect of selenite on the ability of cells to form colonies or to synthesize RNA. We have
found that treatment of HeLa cells with cycloheximide inhibited protein synthesis by >90% but had no effect on the inhibitory
effect of selenite on cell colony formation or RNA synthesis. Since protein synthesis is not necessary for these cytotoxic
effects of selenite they are unlikely to result from an increase in the synthesis of selenoproteins. 相似文献
79.
Structural organization and differential expression of three stilbene synthase genes located on a 13 kb grapevine DNA fragment 总被引:10,自引:0,他引:10
Wilfried Wiese Barbara Vornam Elvira Krause Helmut Kindl 《Plant molecular biology》1994,26(2):667-677
A 13 kb DNA fragment was isolated from a grapevine (Vitis var. Optima) genomic library by hybridizing with elicitor-induced stilbene synthase cDNA as a probe. After fragmentation with Eco RI, subcloning and sequencing, two full-size stilbene synthase genes (Vst1 and Vst2) and the 3 end of a third stilbene synthase gene (Vst3) were located within the 13 kb fragment. Vst1 and Vst2, differing only slightly in the coding region, are distinguished in the intron size and in the structure of the promoter region. The 5 flanking region of gene Vst1 contains a TATAA box at nucleotide –48. The substantial structural differences found for the promoters of the two genes are paralleled by a striking difference in the expression of the two genes in elicitor-treated cells. Moreover, the accumulation upon elicitation of six different stilbene synthase mRNAs was studied and found to differ by two orders of magnitude. 相似文献
80.
Nobuyuki Uozumi Yurie Asano Takeshi Kobayashi 《Plant Cell, Tissue and Organ Culture》1994,36(2):183-190
Adventitious shoot primordia were formed on horseradish hairy root cultured in dark. Plantlet formation frequency from the primordia was higher than that from root fragments. Culture for 26 days provided the adventitious shoot primordia, which had the highest potential for plantlet formation (53% explants at 40 days). Benzyladenine supplementation in the dark caused primordium enlargement, but did not increase the number of primordia formed. After adventitious shoot primordia were encapsulated with calcium alginate, kinetin supplementation (2.0–4.0 M) increased the shoot formation frequency (65–80% explants at 20 days) in the light, but also promoted the undesirable formattion of multiple shoots. Supplementation with naphthaleneacetic acid (0.27–5.4 M) in the calcium alginate beads in light enhanced the root emergence from primordia without inhibition of plantlet formation when the encapsulated beads were put on the agar-medium without naphthaleneacetic acid. 相似文献