Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.4.21.4 (trypsin)
42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The enterotoxic moiety present in the cell-free culture supernatants of Salmonella typhiurium strains (p/536 and p/603) was purified to apparent homogeneity by salt precipitation with ammonium sulphate and successive chromatography through Sephadex G-100 and G-200 columns. It was non-dialysable, heat labile at 90 degrees C and active within pH 6-8. Its activity was completely lost on treatment with trypsin, protease and papain. The enterotoxin appeared to be of high molecular weight (100 kDa) and was highly immunogenic in rabbit. Antigenically, it was not related to cholera toxin, Shiga toxin or the heat labile enterotoxin of Escherichia coli. It did not bind to the GM1 ganglioside. The enterotoxic, delayed permeability and CHO cell elongation activities were attributed to a single protein moiety.
...
PMID:Purification and characterization of enterotoxic moiety present in cell-free culture supernatant of Salmonella typhimurium. 804 72

Saposin B is involved in the hydrolysis of sulfatides, GM1 ganglioside, globotriaosylceramide, and several other sphingolipids and glycerolipids by lysosomal hydrolases. Saposin B is one of four small glycoproteins (saposins) derived from prosaposin. The carbohydrate chain of saposin B was removed and deglycosylated saposin B was characterized and compared with native saposin B. Deglycosylated saposin B stimulated the enzymatic hydrolysis of ganglioside GM1 by acid beta-galactosidase and sulfatide by arylsulfatase A to the same extent as native saposin B. In addition deglycosylated saposin B bound sulfatide and GM1 ganglioside identical to native saposin B. The stability of native saposin B to proteolytic digestion was unchanged by deglycosylation. Neither native saposin B nor deglycosylated saposin B were hydrolyzed by trypsin, endoproteinase Glu-C (V-8), chymotrypsin, or a mixture of acid proteases isolated from human testis. Unlike its effect on metabolic stability, the carbohydrate chain appears to affect folding of saposin B. When native and deglycosylated saposin B were reduced under denaturing conditions and refolded under identical conditions examination of the refolded products indicated that each protein was refolded in a qualitatively different way. A human mutation in saposin B-deficient metachromatic leukodystrophy, in which its glycosylation site is eliminated, has been reported. Our observations suggest that instability of the mutated saposin B is not due to the absence of a protective effect of the carbohydrate chain on proteolysis, but is likely due to aberrant folding resulting from the absence of a carbohydrate chain.
...
PMID:The effect of carbohydrate removal on stability and activity of saposin B. 809 82

In freshly dispersed guinea-pig taenia coli myocytes, beta-adrenergic receptor agonist isoproterenol (ISO, 2-5 microM) increased the open probability (po) of the maxi-K+ channels through the G-protein, G. Subunit B of cholera toxin (0.1 nM) suppressed the ISO-induced increase of po of maxi-Ks+ channels but did not affect that induced by forskolin. Brief (20 min) treatment of the myocytes with GM1 ganglioside (GM1, 0.1-1.0 microM) enhanced the effectiveness of ISO-induced increase of po. This effect was blocked by 0.5% trypsin, which is known to prevent the incorporation of exogeneous GM1 into the membrane. The effect of GM1 was not shared by GM2 and GM3 gangliosides. These results suggest that the membrane-bound endogeneous GM1 may participate in the regulation of cellular response to beta-adrenergic agents.
...
PMID:Enhancement of beta-adrenergic receptor activation of maxi-K+ channels by GM1 ganglioside. 818 85

Anti-gangliotetraosylceramide (anti-asialo GM1) and antiparagloboside monoclonal antibodies (MAbs) were used in immunofluorescence, immunoelectron-microscopic, and in vitro binding inhibition assays to determine whether either of the glycolipids was detectable in the normal cornea, whether levels changed following corneal scarification and either trypsin treatment or incubation in vitro with Pseudomonas aeruginosa, and whether either of the MAbs could competitively inhibit P. aeruginosa binding to cornea. No immunostaining above background for either glycolipid was observed in frozen, unfixed sections or in lightly fixed, K4M-embedded antibody-gold-labeled thin sections of normal cornea. In frozen sections of organ-cultured scarified cornea, no increased immunostaining for anti-asialo GM1 or antiparagloboside reactivity was noted immediately or 60 min after corneal scarification. However, at 60 min after scarification and in vitro incubation of the eye with either trypsin or P. aeruginosa, enhanced immunostaining for both glycolipids was associated with cells within or immediately adjacent to the wound site. Trypsin increased immunoreactivity in the wound site more markedly compared with incubation with P. aeruginosa, but immunostaining was similarly localized with either treatment. No staining above background was seen in control sections. Similarly, with immunoelectron microscopy, increased immunogold-MAb staining for both glycolipids was seen on the plasma membranes of the wound-site cells of eyes incubated with either trypsin or P. aeruginosa compared with controls that were similarly immunostained but with the primary antibody either omitted or substituted with a nonspecific MAb. Competitive binding inhibition assays, in which the bacterial inoculum or the eye in organ culture was incubated with anti-asialo GM1 MAb prior to topical ocular application of the bacteria, showed significantly decreased P. aeruginosa adhesion compared with preparations similarly treated with phosphate-buffered saline or antiparagloboside MAb. These data provide evidence to support the hypothesis that asialo GM1, not paragloboside, serves as a receptor for P. aeruginosa binding to the scarified cornea of the adult mouse and spatially localizes both glycolipids in the wound site.
...
PMID:Evidence for asialo GM1 as a corneal glycolipid receptor for Pseudomonas aeruginosa adhesion. 822 93

Cholera toxin is an ubiquitous activator of intracellular adenylate cyclase and is divided in two major components: A and B. The B-component consists of several subunits that specifically bind to the external cell membrane. The receptor for the toxin, the GM1 ganglioside, is concentrated in nervous tissues. The B subunit of the cholera toxin, conjugated to different molecules (i.e., choleragenoid) is therefore a sensitive anatomical tracer and has been used to detect the presence of GM1 in mammalian tissues. Using choleragenoid, unlabeled and labeled with FITC, we have determined the distribution of the GM1 ganglioside in the vestibular system of the chinchilla. Vestibular tissues were fixed in 4% paraformaldehyde in phosphate buffer, decalcified in 10% EDTA and prepared as either whole-mount, surface-preparations, or for radial cryosections. Positive control tissue consisted of binding to normal brain tissues. Negative controls consisted of several treatments: masking of the GM1 receptors with unlabeled choleragenoid, tissue extraction of GM1 using ethanol, and preabsorbing the choleragenoid with bovine GM1. In addition, to exclude staining of glycoproteins that may have a carbohydrate structure similar to GM1, tissues were digested with trypsin prior to choleragenoid exposure. In the vestibular system, a strongly positive reaction was observed in: the sensory stereocilia and supporting cells of the maculae and cristae, epithelial cells of the planum semilunatum, and polygonal cells of the semicircular canal. Positive but less strong reactivity was observed in the sensory cell body of maculae and cristae, nerve fibers, epithelial cells of utricle and ampulla walls and flattened epithelial cells of the semicircular canals. No reactivity was present in the supporting connective tissue cells and fibrils, blood vessels, gelatinous cupula of the cristae ampullaris and statoconial membranes. Brain tissue showed strong choleragenoid reactivity. The negative controls showed no or greatly reduced reactivity to choleragenoid. Trypsin digestion did not decrease reactivity to choleragenoid.
...
PMID:Localization of the GM1 ganglioside in the vestibular system using cholera toxin. 843 86

Incorporation and metabolism of exogenous GM3 in human myelogenous leukemia HL-60 cells were analyzed using 3H-labeled GM3 ([3H]GM3). [3H]GM3 was rapidly internalized into the cells (trypsin-resistant fraction) 8 times more than the control, 3H-labeled GM1 ([3H]GM1). In addition, not only incorporation but also metabolism of [3H]GM3 was more rapid than [3H]GM1 in HL-60 cells. Moreover, one of the metabolites was found to co-migrate with ceramide in thin-layer chromatography analysis and ceramide formation from exogenous GM3 is more rapid than that from exogenous GM1. These results suggested that there would be some preferential mechanism to produce ceramide from differentiation-inducible GM3 in HL-60 cells rather than from non-inducing GM1.
...
PMID:Rapid internalization of exogenous ganglioside GM3 and its metabolism to ceramide in human myelogenous leukemia HL-60 cells compared with control ganglioside GM1. 900 29

Effects of gangliosides GM1 and GQ1b on cholinergic synaptic functions were investigated using synaptosomes prepared from mouse brain cortices. Treatment of synaptosomes with GM1 and GQ1b increased high K(+)-evoked acetylcholine (ACh) release in a bell-shaped dose-dependent manner. The peaks of the effects were found to be at 1-5 microM for GM1 and 5-10 microM for GQ1b. ACh synthesis and the levels of ACh in synaptosomes were not affected by the ganglioside treatment. Both gangliosides enhanced depolarization-induced influx of calcium ions into synaptosomes. These results indicate that GM1 and GQ1b gangliosides increase evoked ACh release by modulating voltage-dependent calcium channels in the synaptic plasma membranes. The effect of GM1 on calcium ion influx remained after repetitive washings, but was almost completely abolished when the bound GM1 was removed by trypsin. This indicates that the fraction of GM1 which was tightly bound to, but not incorporated in synaptic plasma membranes, is responsible for activating the calcium channels.
...
PMID:Gangliosides enhance KCl-induced Ca2+ influx and acetylcholine release in brain synaptosomes. 924 12

Synaptosomes incorporated mixed brain gangliosides at a rapid initial rate followed by a slower phase of net movement from the protein-associated fraction into the membrane core. The pattern of incorporated gangliosides reflected the pattern available for incorporation. Intact synaptosomes incorporated approximately 100 pmol GM1/mg protein. Synaptosomes preincubated with proteolytic enzymes (trypsin, chymotrypsin, and papain) at different pH values (6.2, 7.4, 7.8) incorporated more exogenous gangliosides than synaptosomes preincubated in buffer alone. This effect was maximal at pH 7.8, though analysis of variance revealed that the proteolytic treatment and pH effects were probably independent processes. Overall uptake of exogenous gangliosides correlated significantly with amount of membrane protein loss, indicating that initial access of exogenous gangliosides to synaptosomal membranes is retarded by cell-surface proteins. These results suggest synaptosomes as a useful alternative to cultured cells for investigating the interaction of gangliosides with other cell surface constituents.
...
PMID:Uptake of exogenous gangliosides by rat brain synaptosomes. 982 Nov 55

To detect HL-60 human promyelocytic leukemia cell proteins involved in the uptake of gangliosides from the culture medium we used photoreactive, 4-azidosalicylic acid (ASA) acylated and radioiodinated (200 Ci/mmole) derivatives of GM3, GD3, GM1, and FucGM1 gangliosides. Gangliosides-ASA, added to the medium at 15-20 nM concentration, followed a similar time course of uptake. After 1 min incubation cell bound gangliosides-ASA could not be removed with trypsin, but only 5-10% remained after incubation with BSA. The proportion of cell bound gangliosides-ASA resistant to BSA treatment increased with time of incubation up to 76% after 20 h. As shown on TLC, GM3- and GD3-ASA were catabolized to LacSph-ASA and ceramide-ASA, while GM1-ASA was hydrolyzed to GM2-ASA. FucGM1-ASA was converted to GM1-ASA very slowly. Upon irradiation with UV lamp, cell bound gangliosides-ASA crosslinked to and photolabeled many proteins but the distribution of radioactivity after SDS/PAGE was very uneven and did not correlate with Coomassie staining. In all experiments the 42 kDa protein bands were most intensely photolabeled. Photolabeling of 42 kDa proteins decreased with time of incubation as compared to lower molecular mass pro teins. With all gangliosides-ASA used similar but not identical protein photolabeling patterns were obtained. Photolabeling patterns with GM3- and GD3-ASA differed from those with GM1- and FucGM1-ASA.
...
PMID:Photochemical labeling of HL-60 cell membrane proteins with radioiodinated, 4-azidosalicylic acid acylated derivatives of gangliosides. 982 71

Photoreactive gangliosides of high specific radioactivity may prove useful for studies on glycosphingolipid functions. We prepared 4-azidosalicylic acid (ASA) acylated derivatives of GM3, GD3, GM1, and FucGM1 gangliosides (gangliosides-ASA). Gangliosides-ASA were characterized by their TLC mobility, UV spectra, carbohydrate composition, and digestion with leech endoceramidase. After radioiodination to about 200 Ci/mmole gangliosides-ASA were used for photochemical labeling of human erythrocytes. Radioiodinated gangliosides-ASA were incorporated into erythrocytes in a time and concentration dependent manner, the kinetics and extent of incorporation being similar for all the gangliosides-ASA used. Radioiodinated gangliosides-ASA incorporated into erythrocytes were resistant to trypsin digestion while treatment with 1% BSA removed about 90% of the label. Incubation with cholera toxin protected radioiodinated GM1-ASA and, to a lesser extent, FucGM1-ASA but not GM3-ASA and GD3-ASA, against removal with BSA. After photolysis about 40-50% of radioactivity was firmly bound to erythrocyte lipids and proteins. The ratio of lipid- to protein-bound radioactivity ranged from 2.2:1 to 3.2:1. Photolabeled proteins were analyzed by SDS/PAGE followed by autoradiography. Band 3 was the most extensively photolabeled protein with all the radioiodinated gangliosides-ASA used. DIDS, an inhibitor of band 3 protein activity, caused reduction in photolabeling of this protein by about 20%.
...
PMID:Photochemical labeling of human erythrocyte membrane proteins with radioiodinated 4-azidosalicylic acid derivatives of G(M3), G(D3), G(M1), and FucG(M1) gangliosides. 982 80


<< Previous 1 2 3 4 5 6 Next >>

---