Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The recently reported phenomenon that red blood cells (RBC) from Alzheimer disease (AD) patients and normal individuals, which have identical electrophoretic mobilities (EPM) in phosphate-buffered saline (PBS), have different EPM in appropriately selected polymer solutions, has been further explored. Of a number of in vitro treatments to which AD and normal RBC were subjected prior to EPM measurements in bottom phase (from a dextran-poly(ethylene glycol) (PEG) aqueous phase system) only trypsin eliminated the difference. Thus, the differential polymer interaction between AD and normal RBC, thought to be the basis for their dissimilar EPM, can be abolished by appropriate proteolytic modification of the cell surfaces and suggests protein as a source of difference. Because young and old RBC from normal individuals, which have the same EPM in PBS, have different EPM in certain polymer solutions, and the RBC from AD patients have been reported to age abnormally, we also compared the young and old RBC subpopulations from these two sources. By the criterion of cell electrophoresis in polymer solutions the differences between AD and normal RBC and between young and old RBC are distinct. The EPM of AD and normal RBC differ in bottom phase or PEG but not in dextran solution; while the EPM of young and old RBC differ predominantly in dextran. We speculate that since the observed difference in EPM of RBC from AD patients and normals depends on protein(s) yet is anticoagulant-related (being obtained only when blood is collected in citrate or oxalate) it might be the result of an interaction (Ca(2+)-mediated?) between the surfaces of these cells and protein component(s) of their respective, compositionally differing sera.
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PMID:Differential electrophoretic behavior in aqueous polymer solutions of red blood cells from Alzheimer patients and from normal individuals. 753 1

The interaction between bovine lactoferrin (bLf) and ascorbate (Asc) was investigated through malondialdehyde (MAD) formation in a solution containing DNA, bleomycin (BLM), and Fe2+ or Asc. The inhibition by bLf on MDA formation in the presence of Asc was not changed even by adding carbonate or oxalate ions to the solution. The percentage inhibition by the hydrolysates of bLf treated with pepsin, trypsin, and both enzymes on MDA formation was almost the same as that by the untreated bLf in the presence of Asc. The inhibition of MDA formation also occurred with the filtrate obtained from a solution containing bLf and Asc, but not with that from a solution of bovine serum albumin and Asc. The interaction of bLf and Asc was observed by gel filtration in a Sephadex G75 column. The binding amount of Asc was estimated to be 87 mol per mole of bLf.
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PMID:Interaction of lactoferrin with ascorbate and the relationship with bleomycin-dependent DNA damage. 753 54

This study was conducted to determine whether a factor responsible for reduced nicotinamide adenine dinucleotide phosphate (NADPH)-supported lipid peroxidation in rat liver microsomes is involved in iron reduction by cooperation with NADPH-cytochrome P450 reductase. Under anaerobic conditions, NADPH-dependent reduction of ferric pyrophosphate in microsomes was not dependent on cytochrome P450 levels and was not inhibited by carbon monoxide (CO). All of the iron complexes with chelators such as adenosine 5'-diphosphate, pyrophosphate, nitrilotriacetate, oxalate or citrate were reduced in microsomes, although in the reconstituted system containing purified NADPH-cytochrome P450 reductase little or no iron reduction was found. A cytochrome P450-free fraction from a cholate-solubilized preparation of microsomes after passage through a laurate sepharose column was required for reduction of iron pyrophosphate in the reconstituted system leading to lipid peroxidation. The iron reduction was not inhibited by CO and was destroyed by heat treatment or trypsin digestion of the fraction. All iron complexes were reduced in the presence of the fraction, using a reducing equivalent of NADPH via NADPH-cytochrome P450 reductase. The results indicate that a heat-labile component, which is probably a protein distinct from cytochrome P450, is associated with iron reduction responsible for lipid peroxidation in microsomes.
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PMID:A microsomal membrane component associated with iron reduction in NADPH-supported lipid peroxidation. 776 Jun 89

We investigated the effects of the nonionic detergent octaethylene glycol monododecyl ether (C12E8) on the sarcoplasmic reticulum calcium pump in cardiac microsomes in view of its specific effects on different ATP-accelerated steps in the catalytic cycle of the Ca-ATPase in leaky fast skeletal muscle microsomes. At low concentrations of MgATP2- (< 2.5 microM), a nonsolubilizing concentration of added C12E8 (15 microM) increased apparent Vmax(MgATP) of oxalate-facilitated calcium uptake associated with MgATP2- binding to the high affinity catalytic site. An ATP induced acceleration of calcium uptake, attributable to regulatory nucleotide binding, was seen between 2 and 3 microM MgATP2- in both C12E8-treated and control microsomes. These effects of C12E8 are similar to those seen previously with trypsin treatment of microsomes [Lu, Y.-Z., Xu, Z.-C., & Kirchberger, M.A. (1993) Biochemistry 32, 3105-3111]. However, at a saturating Ca2+ between 3 and 10 microM MgATP2-, C12E8 produced a greater reduction in the magnitude of the ATP-induced acceleration of calcium uptake seen with trypsin. At 1 mM MgATP2-, C12E8 and trypsin as well as protein kinase A-catalyzed microsomal phosphorylation all increased the Ca2+ affinity of the pump, but only the latter two treatments significantly increased apparent Vmax(Ca). In fact in trypsin-treated and phosphorylated microsomes, C12E8 reduced Vmax(Ca) to close to the control values; it reduced Vmax(Ca) only slightly in control microsomes. Under our experimental conditions, comparable effects of 15 microM C12E8 on calcium uptake were absent in fast skeletal muscle microsomes, which lack phospholamban.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effects of a nonionic detergent on calcium uptake by cardiac microsomes. 817 81

Uronic-acid-rich protein (UAP) is a urinary glycoprotein that inhibits calcium oxalate crystallization in vitro. It shows a structural similarity to bikunin, a component of inter-alpha-inhibitor (IalphaI) known for its inhibition of the action of many serine proteinases like trypsin and chymotrypsin. To clarify the relationship between these macromolecules, UAP, IalphaI, urinary bikunin, and plasma bikunin were purified and studied. Their calcium oxalate crystallization inhibitory activity was assayed before and after treatment with chondroitinase AC and pronase. Their molecular mass was determined by using SDS/PAGE before and after these treatments. Polyclonal bikunin antibody was used on Western blots for immunological identification. The partial amino acid sequence of UAP before and after chondroitinase treatment was determined. Also, the antitryptic activity of UAP was measured and compared to that of bikunin, which is responsible for the antiprotease activity of IalphaI. UAP exhibited a strong calcium oxalate crystallization inhibitory activity. IalphaI and both bikunins were less inhibitory. Chondroitinase AC had no effect on inhibitory activity of these proteins even when their molecular mass changed. However, after pronase treatment, the inhibitory activity of both bikunins and UAP was completely destroyed. The antitryptic activity of UAP was found to be 0.78 U/mg which is lower than that of bikunin which is about 1.9 U/mg. On Western blotting, bikunin antibody immunoreacted with UAP and both urinary and plasma bikunins. Partial amino acid sequence confirmed the identity of UAP as urinary bikunin.
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PMID:Identification of uronic-acid-rich protein as urinary bikunin, the light chain of inter-alpha-inhibitor. 866 22

As the search for alternative sources of food to alleviate hunger continues, this study was undertaken to determine the biological value in growing rats (BV) of proteins of some lesser known tropical seeds gathered in Nigeria. Antinutritional factors (trypsin inhibitors, phytic acid, oxalate, tannin, alkaloids) and amino acid compositions were also determined, and protein digestibility-corrected amino acid score (PDCAAS) was calculated using the amino acid requirement pattern of the preschool child and individual seed-specific correction factors for crude protein. A rat growth and balance study was conducted to determine digestibility, nitrogen-, and energy balance by feeding as the only unsupplemented protein source milled and heat-treated seeds of Adansonia digitata (Bombacaceae) and Prosopis africana, Lonchocarpus sericeus, Enterolobium cyclocarpium, Sesbania pachycarpa and Pterocarpus osun (Leguminosae) in comparison to casein fortified with methionine (control). Diets containing P. africana and L. sericeus seeds caused poor feed intake and weight loss in rats and were excluded from the nitrogen-balance test. Among the seed samples, S. pachycarpa followed by A. digitata showed the most advantageous nutritional quality [amino acid composition, digestibility, BV and net protein utilization (NPU)]. True digestibility was 82.9 and 74.5 vs. 98.5, BV was 64.6 and 70.0 vs. 90.4, and NPU was 53.5 and 52.1 vs. 89.0 for S. pachycarpa and A. digitata vs. casein (control), respectively. In terms of PDCAAS, lysine was the first limiting amino acid for S. pachycarpa (88%) and for A. digitata (58%). The PDCAAS of all essential amino acids was below 100% for E. cyclocarpium (e.g., cysteine + methionine: 37%) and for P. africana (e.g., threonine: 46%, except valine and a very high content of cysteine and methionine). In conclusion, all seeds tested in the rat balance trial were of inferior quality compared to casein. Before these tropical seeds could be used as food components or feed supplements, safety studies and proper processing to remove antinutritional factors and possible toxic constituents were required.
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PMID:Low nutritional quality of unconventional tropical crop seeds in rats. 980 58

Inter-alpha-inhibitor (I alpha I) is a serine protease inhibitor present in human plasma. It has a molecular weight of about 220 kDa which encompasses 3 chains including two heavy chains and one light chain. The light chain, known as bikunin, is responsible for the antitryptic activity of I alpha I in the inhibition of various enzymes, such as trypsin and chymotrypsin. Under physiologic or certain pathologic circumstances, several macromolecules related to I alpha I appear in plasma and urine. However, the physiologic role of I alpha I remains unclear. As far as urolithiasis is concerned, two urinary macromolecules related to I alpha I have been isolated and shown to be potent inhibitors of calcium oxalate formation. One of these inhibitors, uronic-acid-rich protein (UAP), has been identified and well characterized. The sequence of the first 18 amino acid residues of UAP is identical with that of bikunin. Furthermore, the immunoreaction between UAP and I alpha I antibody using immunoblot analysis was positive. UAP isolated from the urine of stone formers exhibited less inhibitory activity towards calcium oxalate crystallization than that derived from the urine of healthy subjects. This suggests a structural abnormality of the inhibitor obtained from stone patients. The organic matrix extracted from kidney stones contained a protein antigenically related to I alpha I. We conclude that UAP is a member of I alpha I family taking part in inhibiting calcium oxalate crystallization, and modulating the formation of stones in the urinary tract.
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PMID:Inter-alpha-inhibitor: a protein family involved in the inhibition of calcium oxalate crystallization. 981 21

Two proteins of 17 and 24 kDa, respectively, which were immunologically related to bikunin, were purified from urine of healthy men, using in the last step a trypsin CNBr-sepharose affinity column. These proteins strongly inhibited calcium oxalate (CaOx) crystallization in two in vitro models. In the first model, the presence of 8 microg/ml protein in a medium containing 0.76 mM CaCl2 (with 45Ca) and 0.76 mM ammonium oxalate inhibited the crystallization process by 80%, as estimated by supernatant radioactivity after 60 min of incubation. A similar inhibition was observed in the second turbidimetric model, where the CaOx crystallization kinetics were followed for 10 min at 620 nm in a medium containing 4 mM CaCl2 and 0.5 mM Na2Ox. These proteins were used as standard protein for the development of an enzyme-linked immunosorbent assay (ELISA) in urine. Mean (+/-SEM) urinary bikunin concentration in 18 healthy subjects was 5.01 +/- 0.91 microg/ml. This was a concentration range of strong inhibitory activity in vitro. Bikunin values were nearly 50% lower (2.54 +/- 0.42 microg/ml, P=0.007) in 31 CaOx renal stone formers (having weddelite crystals in their first morning urine) than in the healthy volunteers. A correlation was found between urinary bikunin and alpha-1 microglobulin concentrations in the control group (y=0.73x + 1.09, r2=0.8) while no such correlation existed in the lithiasis group. In conclusion, bikunin exerts a strong inhibitory action of CaOx crystallization in vitro. Its involvement in urinary CaOx crystallization of stone formers is highly probable, based on the significant decrease in its urinary concentration in the majority of stone formers studied.
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PMID:Inhibitory effect of bikunin on calcium oxalate crystallization in vitro and urinary bikunin decrease in renal stone formers. 1009 56

The effects of heat treatments on the proximate composition, energy content, and levels of some antinutritional factors in brown and marble-colored African yam bean (AYB) seed flours were investigated. In raw brown and marble-colored AYB seed flours; moisture content, dry matter, crude protein, crude fat, ash, total carbohydrate and caloric value did not differ significantly at the 5% level. Autoclaving and cooking slightly increased the moisture level. Crude protein, crude fat, and ash contents were decreased by autoclaving and were further decreased by cooking. The decrease was not, however, considerable for the AYB that is not eaten raw and whose full nutritional potential as a legume can be derived only when heat treated, as previous reports have indicated for legume seeds. The levels of the toxicants were generally higher in the raw brown AYB compared to the marble-colored, and were generally reduced by both autoclaving and cooking. In the most commonly available and consumed marble-colored AYB, autoclaving at 121 degrees C, 15 psi for 20 min decreased cyanogenic glycosides by 46%, oxalate by 48.9%, tannin by 15.0%, saponin by 14.8% and trypsin inhibitors by 61.3% while cooking for 3.5 hours in tap water decreased these toxic factors by 66.5%, 70.3%, 72.2%, 48.7%, and 86.0%, respectively. The results indicate that for raw samples, varietal difference did not significantly affect nutrient composition though the toxicants were generally higher in the brown AYB than the marble-colored. Autoclaving decreased both nutrient value and the level of toxicants in the two seed types; values were further reduced by cooking. Of the toxicants, trypsin inhibitor was found to be the most heat-labile and of the heat treatment methods, cooking to tenderness is recommendable.
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PMID:Effect of heat treatment on the proximate composition, energy values, and levels of some toxicants in African yam bean (Sphenostylis stenocarpa) seed varieties. 1260 31

1. Crystallized soy bean trypsin inhibitor, at a concentration of 100 microg./ml., suppressed the production of thrombin from a mixture of prothrombin and blood thrombokinase. The experiment was performed in the presence of 0.011 M oxalate, in order to minimize the possibility of participation by accessory factors which require ionic calcium. The results are in accord with the view that thrombokinase is a trypsin-like enzyme. 2. When a solution of blood thrombokinase was centrifuged at 85,000 g for 120 minutes, almost all the activity remained in the supernate. This supernate activated the supernate from a prothrombin solution which had been similarly centrifuged. The activation of prothrombin by thrombokinase can proceed in the absence of material completely sedimentable in 120 minutes at 85,000 g. 3. An "accelerator" reagent was prepared by treating bovine serum with barium carbonate, and then passing the serum through a column of diatomaceous earth. This "accelerator" was used together with prothrombin, blood thrombokinase, Howell's cephalin, and calcium chloride to compose a five-reagent thrombin-producing system. In this system, no thrombin was produced without thrombokinase. On the other hand, thrombin was produced from prothrombin and thrombokinase, even when all the other reagents were omitted. When calcium was omitted, thrombokinase was able to function; but cephalin and the "accelerator" reagent were ineffective. 4. Quantitative tests indicated that the "accelerator" reagent exerted an effect distinct from those of thrombokinase and cephalin. However, it is not certain whether the "accelerator" reagent functioned as an accessory factor, as a potential source of more thrombokinase, or both. In the experiments reported, thrombokinase was primary to, or necessary for, the effect of "accelerator." 5. The effectiveness of thrombokinase was multiplied a hundred times or more, when complemented by calcium, cephalin, and "accelerator" reagent. Ionic calcium was a necessary component of this complementing system. This may help to explain why removal of calcium ions keeps blood fluid, even though thrombokinase, by itself, is little influenced either by calcium ions or by oxalate.
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PMID:Effect of blood thrombokinase, as influenced by soy bean trypsin inhibitor, ultracentrifugation, and accessory factors. 1324 62


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