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1.
Identification and partial characterization of the human erythrocyte membrane component(s) that express the antigens of the LW blood-group system. 总被引:7,自引:2,他引:5
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G Mallinson P G Martin D J Anstee M J Tanner A H Merry D Tills H H Sonneborn 《The Biochemical journal》1986,234(3):649-652
Rhnull human erythrocytes lack the antigens of the Rhesus blood-group system, have an abnormal shape, have an increased osmotic fragility, and are associated with mild chronic haemolytic anaemia. Rhnull erythrocytes also lack all antigens of the LW blood-group system, but the functional significance of this deficiency is unknown. We have identified, by immunoblotting with two mouse monoclonal antibodies (BS46 and BS56), the LW-active component(s) in normal human erythrocytes as a broad band of Mr 37 000-47 000 on SDS/polyacrylamide-gel electrophoresis. Treatment of intact human erythrocytes with endoglycosidase F preparation destroyed the epitopes recognized by antibodies BS46 and BS56, suggesting that one or more N-glycosidically linked oligosaccharides are required for the formation of the LW antigens. Estimation of the number of LW antigen sites per erythrocyte by using radioiodinated purified antibody BS46 gave average values of 4400 molecules/cell for Rh(D)-positive adult erythrocytes and 2835 molecules/cell for Rh(D)-negative adult erythrocytes. Like the Rh(D) polypeptide, the LW polypeptide(s) is (are) associated with the cytoskeleton of normal erythrocytes. These results suggest the possibility that the absence of the LW polypeptide may also contribute to the functional and/or morphological abnormalities of Rhnull erythrocytes. 相似文献
2.
Immunochemical studies on the specificity of soybean agglutinin 总被引:9,自引:0,他引:9
The specificity of the purified soybean agglutinin has been studied immunochemically by quantitative precipitin and quantitative precipitin inhibition assays. The lectin is precipitated by human A and Lea blood-group substance, by the products of the second, third, fourth, and fifth stages of periodate oxidation of a human H blood-group substance (JS), and by precursor blood-group substances, as well as by a pig-submaxillary mucin having blood-group A activity, by partially hydrolyzed blood-group B substances (Pl fraction), and by group C streptococcal polysaccharide. The activity is attributable to terminal α-linked 2-acetamido-2-deoxy-d-galactopyranosyl or to α- or β-d-galactopyranosyl residues. The lectin did not precipitate with human blood-group H substances, with the product of the first stage of periodate oxidation (JS), with streptococcal group A polysaccharide, or with pig-submaxillary mucin devoid of blood-group A activity, and is poorly precipitated by blood-group B substances. Inhibition of precipitation with various monosaccharides indicated that the lectin is strongly specific for 2-acetamido-2-deoxy-d-galactose and for its oligosaccharides, and to a lesser extent for d-galactose and its oligosaccharides; the α-glycosides of both sugars were slightly more reactive than the β-glycosides of 2-acetamido-2-deoxy-d-galactose, and both α- and β-glycosides were more active than the free monosaccharides. Aromatic α- and β-glycosides of 2-acetamido-2-deoxy-d-galactose and d-galactose were better inhibitors than the corresponding methyl or ethyl compounds. The blood-group A trisaccharide α-d-GalNAcp-(1→3)-β-d-Galp-(1→3)-d-GlcNAc was more active than the disaccharide lectins by the use of precipitation with polysaccharides, as well as inhibition reactions, is essential to the understanding of their reactivity with cell-surface receptors. 相似文献
3.
Than NG Romero R Meiri H Erez O Xu Y Tarquini F Barna L Szilagyi A Ackerman R Sammar M Fule T Karaszi K Kovalszky I Dong Z Kim CJ Zavodszky P Papp Z Gonen R 《PloS one》2011,6(7):e21564
Background
Placental Protein 13 (PP13), an early biomarker of preeclampsia, is a placenta-specific galectin that binds beta-galactosides, building-blocks of ABO blood-group antigens, possibly affecting its bioavailability in blood.Methods and Findings
We studied PP13-binding to erythrocytes, maternal blood-group effect on serum PP13 and its performance as a predictor of preeclampsia and intrauterine growth restriction (IUGR). Datasets of maternal serum PP13 in Caucasian (n = 1078) and Hispanic (n = 242) women were analyzed according to blood groups. In vivo, in vitro and in silico PP13-binding to ABO blood-group antigens and erythrocytes were studied by PP13-immunostainings of placental tissue-microarrays, flow-cytometry of erythrocyte-bound PP13, and model-building of PP13 - blood-group H antigen complex, respectively. Women with blood group AB had the lowest serum PP13 in the first trimester, while those with blood group B had the highest PP13 throughout pregnancy. In accordance, PP13-binding was the strongest to blood-group AB erythrocytes and weakest to blood-group B erythrocytes. PP13-staining of maternal and fetal erythrocytes was revealed, and a plausible molecular model of PP13 complexed with blood-group H antigen was built. Adjustment of PP13 MoMs to maternal ABO blood group improved the prediction accuracy of first trimester maternal serum PP13 MoMs for preeclampsia and IUGR.Conclusions
ABO blood group can alter PP13-bioavailability in blood, and it may also be a key determinant for other lectins'' bioavailability in the circulation. The adjustment of PP13 MoMs to ABO blood group improves the predictive accuracy of this test. 相似文献4.
Three isomeric ceramide tetrasaccharides — P blood-group active globoside, lacto-N-neotetraosyl ceramide as ABH blood-group precursor, both isolated from human erythrocytes and “asiologanglioside” from human brain as reference standard — and two ceramide pentasaccharides — H blood-group active glycosphingolipid, obtained from blood-group B active ceramide hexasaccharide of human B erythrocytes after α-galactosidase treatment and ceramide pentasaccharide from rabbit erythrocytes with B-like blood-group activity — were investigated by mass spectrometry after permethylation. The carbohydrate moiety exhibits differences not only concerning the sugar sequence but also with regard to the position of some glycosidie linkages: Oligosaccharides containing N-acetylhexosamine substituted at position 4 produce spectra that are distinctly different from those containing C-3 substituted N-acetylhexosamines, thus allowing the differentiation between type 1 and type 2 carbohydrate chains. Moreover, oligosaccharide ions with a hexose at the cleavage site exhibit a fragmentation pattern different from those with a N-acetylhexosamine at the “reducing terminal”. The intensity ratio between parent ion and parent ion — 32 mass units is Q ? 3 in the first case, whereas in the latter case Q is <1. The Q-values are given for 14 oligosaccharide ions. Differences in the composition of the ceramide residues can also be deduced from the mass spectra. 相似文献
5.
The structure of a complex fucolipid from hog gastric mucosa containing twenty sugar residues and exhibiting blood-group (A + H) activity has been investigated. Based on the results of immunological assays, partial acid hydrolysis, sequential degradation with specific exoglycosidases, oxidation with periodate and chromium trioxide, and permethylation analysis, we suggest that the carbohydrate chain of this fucolipid contains four termini. One of the termini bears beta Gal1 leads to 4 beta GlcNAc disaccharide, two bear blood-group A determinant and one bears H determinant. Two of the branches, terminated by beta Gal1 leads to 4 beta GlcNAc and blood-group A determinant, and two terminated by blood-group A and H determinants, are linked through beta Gal1 leads to 4 beta GlcNAc1 leads to 3/6 and beta Gal1 leads to 4 beta GlcNAc1 leads to 4 beta GlcNAc1 leads to 3/6 to the galactose residue adjacent to glucosylceramide core. 相似文献
6.
More than ten new types of gangliosides, in addition to haematoside and sialosylparagloboside, were isolated from human erythrocyte membranes. These were separated by successive chromatographies on DEAE-Sephadex, on porous silica-gel columns and on thin-layer silica gel as acetylated compounds. Highly potent blood-group-Ii and moderate blood-group-H activities were demonstrated in some of the ganglioside fractions. The gangliosides incorporated into cholesterol/phosphatidylcholine liposomes stoicheiometrically inhibited binding of anti-(blood-group I and i) antibodies to a radioiodinated blood-group-Ii-active glycoprotein. The fraction with the highest blood-group-I-activity, I(g) fraction, behaved like sialosyl-deca- to -dodeca-glycosylceramides on t.l.c. Certain blood-group-I and most of the -i determinants were in partially or completely cryptic form and could be unmasked by sialidase treatment. Thus the I and i antigens, which are known to occur on internal structures of blood-group-ABH-active glycoproteins in secretions, also occur in the interior of the carbohydrate chains of erythrocyte gangliosides. 相似文献
7.
The specificity of Bandeiraea simplicifolia lectin I (BS I) has been studied by competitive-binding assays (CBA) using tritium-labeled human B and hog A substances. Blood-group B substances isolated from horse gastric mucosae and from human ovarian-cyst fluids were much better inhibitors of binding of tritiated blood-group B substance to insoluble BS I-Sepharose 2B than were human blood-group A substances from saliva and ovarian-cyst fluid. A and B active blood-group substances showed the same range of potency in inhibiting binding of tritium-labeled hog A substance to BS I-Sepharose 2B. CBA with BS I-Sepharose 2B, labeled human blood-group B substance, and human blood-group A and B active aligosaccharides separated the haptens into two groups differing in slope. Group 1, containing methyl alpha-D-GalNAcp, D-GalNAcp, and an A active pentasaccharide ARL 0.52, with 3, 19, and 25 nmol respectively needed for 50% inhibition of binding, has a lower slope than group 2, which contains alpha-D-GalNAcp-(1 leads to 3)-2-acetamido-2-deoxy-D-galactitol and p-nitrophenyl alpha-D-GalNAcp, with 3 nmol of each required for 50% inhibition of binding, as well as ten glycosides with terminal, nonreducing, alpha-linked D-Galp. The most potent inhibitors of this group were p-nitrophenyl alpha-D-Galp, alpha-D-Galp-(1 leads to 3)-D-Galp, alpha-D-Galp-(1 leads to 6)-D-Glcp, and methyl alpha-D-Galp, with 5, 7.4, 9.6, and 11 nmol respectively needed to inhibit binding by 50%. The difference in slopes was explainable in terms of a recent finding that BS I exists as a mixture of five isolectins composed of two subunits having different specificities; subunit A is most specific for alpha-linked, terminal, nonreducing D-GalNAcp, but it also reacts with alpha-linked, terminal, nonreducing D-Galp, whereas subunit B tends to be more specific for terminal, nonreducing, alpha-linked D-Galp. 相似文献
8.
Carboxypeptidase Y digestion of band 3, the anion transport protein of human erythrocyte membranes 总被引:1,自引:0,他引:1
The exposure of the carboxyl-terminal of the Band 3 protein of human erythrocyte membranes in intact cells and membrane preparations to proteolytic digestion was determined. Carboxypeptidase Y digestion of purified Band 3 in the presence of non-ionic detergent released amino acids from the carboxyl-terminal of Band 3. The release of amino acids was very pH dependent, digestion being most extensive at pH 3, with limited digestion at pH 6 or above. The 55,000 dalton carboxyl-terminal fragment of Band 3, generated by mild trypsin digestion of ghost membranes, had the same carboxyl-terminal sequence as intact Band 3, based on carboxypeptidase Y digestion. Treatment of intact cells with trypsin or carboxypeptidase Y did not release any amino acids from the carboxyl-terminal of Band 3. In contrast, carboxypeptidase Y readily digested the carboxyl-terminal of Band 3 in ghosts that were stripped of extrinsic membrane proteins by alkali or high salt. This was shown by a decrease in the molecular weight of a carboxyl-terminal fragment of Band 3 after carboxypeptidase Y digestion of stripped ghost membranes. No such decrease was observed after carboxypeptidase Y treatment of intact cells. In addition, Band 3 purified from carboxypeptidase Y-treated stripped ghost membranes had a different carboxyl-terminal sequence from intact Band 3. Cleavage of the carboxyl-terminal of Band 3 was also observed when non-stripped ghosts or inside-out vesicles were treated with carboxypeptidase Y. However, the digestion was less extensive. These results suggest that the carboxyl-terminal of Band 3 may be protected from digestion by its association with extrinsic membrane proteins. We conclude, therefore, that the carboxyl-terminal of Band 3 is located on the cytoplasmic side of the red cell membrane. Since the amino-terminal of Band 3 is also located on the cytoplasmic side of the erythrocyte membrane, the Band 3 polypeptide crosses the membrane an even number of times. A model for the folding of Band 3 in the erythrocyte membrane is presented. 相似文献
9.
The human erythrocyte anion exchange protein, Band 3, was reacted with N-ethylmaleimide (NEM) in cells to a stoichiometry of 5.3 mol NEM per mol Band 3, indicating that all NEM-reactive cysteines in Band 3 were labeled. Quantitatively NEM-blocked Band 3 was still able to bind to and be eluted by reducing agents from a mercurial affinity resin, [p-(chloromercuribenzamido)ethylene]amino-Sepharose. Reaction of NEM-blocked Band 3 with p-chloromercuribenzoate (pCMB) did not prevent binding to the resin due to exchange of pCMB for the immobilized mercurial. pCMB has been reported to inhibit water and urea permeation across the red cell membrane, and this has been attributed to reaction with a NEM-reactive sulfhydryl in Band 3. The interaction of Band 3 with the immobilized ligand directly demonstrates the reaction of NEM-blocked Band 3 with a mercurial and indicates that the NEM-unreactive, pCMB-reactive sulfhydryl residue is accessible to within approximately equal to 12 A (the distance from the solid support to the Hg) of the surface of the solubilized Band 3 protein. 相似文献
10.
Characterization of matrix-bound Band 3, the anion transport protein from human erythrocyte membranes 总被引:4,自引:0,他引:4
Band 3 (Mr = 95,000), the anion transport protein of human erythrocyte membranes exists primarily as a dimer in solutions of nonionic detergents such as octaethylene glycol mono-n-dodecyl ether (C12E8). The role of the oligomeric structure of Band 3 in the binding of [14C]4-benzamido-4'-aminostilbene-2,2'-disulfonate (BADS), an inhibitor of anion transport (Ki = 1-2 microM), was studied by characterizing the interaction of BADS with dimers and monomers of Band 3 covalently attached to p-mercuribenzoate-Sepharose 4B. BADS bound to matrix-bound Band 3 dimers with an affinity of approximately 3 microM at a stoichiometry of 1 BADS molecule/Band 3 monomer, in agreement with the BADS binding characteristic of Band 3 in the membrane and in solutions of C12E8. Band 3 dimers could be attached to the matrix via one subunit by limiting the amount of p-chloromercuribenzoate on the Sepharose bead. Matrix-bound monomers were formed by dissociation of the dimers with dodecyl sulfate or guanidine hydrochloride. Complete removal of the denaturants allowed formation of refolded Band 3 monomers since the matrix-bound subunits could not reassociate. These refolded Band 3 monomers were unable to bind BADS. Release of the monomers from the matrix with 2-mercaptoethanol allowed reformation of dimers with recovery of the BADS binding sites. These results suggest that the dimeric structure of Band 3 is required for BADS binding and that the BADS binding sites may be at the interface between the two halves of the Band 3 dimer. 相似文献
11.
《Revue fran?aise de transfusion et immuno-hématologie》1980,23(5):545-552
The oligosaccharide structures specifying the blood-group ABH determinants occur in the human erythrocyte membrane in different classes of compounds. The majority occur in a novel class of complex carbohydrate chains called the polyglycosyl chains. They are bound by an alkali-stable bond to glycoproteins (band 3, band 4.5) and occur also in glycolipids. Conventional glycosphingolipids as well as alkalilabile carboyhydrate chains of glycoproteins (in the PAS-bands) are also carriers of the blood-group determinants. 相似文献
12.
Immunochemical studies on the specificity of the peanut (Arachis hypogaea) agglutinin. 总被引:6,自引:0,他引:6
The specificity of purified, peanut agglutinin has been studied immunochemically by quantitative precipitin and inhibition assays. The lectin showed substantial differences in precipitating with blood-group substances of the same specificity. Of the B substances tested, horse 4 25% completely precipitated the lectin, Beach phenol insoluble failed to interact, and PM phenol insoluble gave an intermediate reaction. The lectin did not precipitate with A1 substances, with hog gastric mucin A + H substance, or with A2 substance WG phenol insoluble. Another A2 substance, cyst 14 phenol insoluble, precipitated approximately 2/3 of the lectin. Of the H substances, Tighe phenol insoluble was inactive, JS phenol insoluble precipitated poorly, and morgan standard H precipitated about 80% of the lectin. However, first stage of Smith degradation, as well as Pl fractions obtained by mild acid hydrolysis of blood-group substances, gave products which precipitated strongly. The lectin was also completely precipitated by all precursor blood-group substances, as well as by cows 21 and 26, all having strong I-Ma, I-Ort, I-Step, and I-Da activities. Cow 18, which does not possess significant blood-group I activity, precipitated very slightly. Fractions of blood-group substances N-1 (Lea) and Tij (B) obtained by precipitation from 90 percent phenol at higher concentrations of ethanol interacted better with peanut agglutinin. These differences in activity are ascribable to a heterogeneity resulting from incomplete biosynthesis of carbohydrate side-chains of blood-group substances, particularly resulting in variations in the numbers of DGalbeta1 leads to 3DGalNAc or DGalbeta1 leads to 4DGlcNAc determinants. The agglutinin reacted with the hydatid cyst P1 glycoprotein, as well as with the previously studied antifreeze and sialic acid-free alpha1 acid glycoproteins, but not with pneumococcus type XIV polysaccharide. Inhibition of precipitation showed the lectin to be most specific for the disaccharide DGalbeta1 leads to 3DGalNAc, which is 14, 55, and 90 times as active as DGalbeta1 leads to 4DGlcNAc, DGal, and DGalbeta1 leads to 3DGlcNAc, respectively. DGalbeta1 leads to 3N-acetyl-D-galactosaminitol has approximately 1/25th the activity of DGalbeta1 leads to 3DGalNAc. Substitutions of DGlcNAc or LFuc on the DGal of active inhibitors completely blocked the activity, in line with the assumption that the combining site of the peanut lectin is a partial cavity. The oligosaccharides DGalbeta1 leads to 4DGlcNAcbeta1 leads to 6-hexane-1,2,4,5,6-pentol(s) and DGalbeta1 leads to 3[DGalbeta1 leads to 4DGlcNAcbeta1 leads to 6]N-acetyl-D-galactosaminitol showed the same inhibitory activity as DGalbeta1 leads to 4DGlcNAc, suggesting that the combining site of the peanut agglutinin may not be complementary to more than a disaccharide... 相似文献
13.
Phosphodiesterase-5 (PDE5) is a dimer containing a cGMP-specific catalytic domain and an allosteric cGMP-binding subdomain (GAF A) on each subunit. PDE5 exhibits three conformational forms that can be separated by Native PAGE and are denoted as Bands 1, 2, and 3 in decreasing order of mobility. A preparation comprised mainly of Band 2 PDE5 was partially converted to Band 3 PDE5 by 1 h incubation with cGMP or the PDE5-specific inhibitors sildenafil, vardenafil, or tadalafil, but not with cAMP, milrinone (PDE3-specific), or rolipram (PDE4-specific). Band 2 PDE5 was converted almost entirely to Band 3 PDE5 by overnight incubation with sildenafil at 30 °C. This time-dependent conversion was accompanied by a 7-fold increase in allosteric cGMP-binding activity, suggesting that Band 3 PDE5 is a much more active form than Band 2 PDE5 for allosteric cGMP binding. Conversion of Band 2 PDE5 to Band 3 PDE5 occurred faster by pre-incubation with cGMP, which binds to both the allosteric and catalytic sites of PDE5, than with catalytic site-specific sildenafil. Overnight incubation of a Band 2/Band 3 PDE5 mixture with EDTA caused time-dependent conversion to Band 1 PDE5 (apoenzyme), and this conversion was accompanied by a 50% loss in cGMP-binding activity. After incubation with EDTA, addition of Mn++ or Mg++ caused reversion of Band 1 to a Band 2/Band 3 PDE5 mixture in which Band 3 PDE5 predominated. This reversion was accompanied by a 3-fold increase in allosteric cGMP-binding activity. The combination of results implied that physiological conversion of Band 2 to Band 3 PDE5 by cGMP and/or divalent metal ion occupancy of the catalytic domain would increase allosteric cGMP binding to the enzyme. This conversion would produce a greater negative feedback effect on cGMP action by increasing sequestration of cGMP at the allosteric cGMP-binding site of PDE5 and by increasing cGMP degradation at the catalytic site of the enzyme. This conversion would also increase PDE5 inhibitor binding to the enzyme. 相似文献
14.
R J Howard P M Smith G F Mitchell 《The Australian journal of experimental biology and medical science》1979,57(4):355-368
Optimal conditions were established for specific labelling of the surface proteins of mouse erythrocytes using lactoperoxidase-catalyzed radioiodination. The levels of H2O2 and I-, and cell concentrations required for restriction of haemoglobin labelling to less than 5% of the total 125I-protein, were different for radioiodination employing direct H2O2 addition or generation of H2O2 with glucose oxidase plus glucose. Preparation of mouse erythrocyte ghosts by hypotonic lysis caused loss of some minor labelled proteins present on intact cells and shifts to lower molecular weights of others. It is therefore important to solubilize labelled cells directly in electrophoresis buffer to avoid artifactual degradation of labelled proteins. The extent of labelling internal cell proteins was measured by a procedure suitable for the comparison of a large number of samples: solubilized radioiodinated erythrocytes were electrophoresed on 14% acrylamide gels and the radioactivity determined in the haemoglobin band which migrates separately from other proteins. The major labelled protein on the mouse erythrocytes had an apparent molecular weight of 92,000, and may be analogous to Band 3 of the human erythrocyte. 相似文献
15.
Naturally occurring IgG autoantibody against Band 3 glycoprotein of human erythrocyte membrane (anti-Band 3) recognizes the erythrocytes modified with oxidizing or SH-blocking agents as well as senescent erythrocytes. Location of the antigenic determinants of Band 3 this autoantibody recognizes was investigated by competitive inhibition studies of the antibody binding to the modified cells. Autologous IgG binds to the modified erythrocytes, and purified Band 3 totally inhibits the binding. This inhibitory activity of Band 3 was not affected by digestion of Band 3 with various proteases. Treatment of Band 3 with endo-beta-galactosidase that destroys the poly-N-acetyllactosaminyl sugar chain of Band 3 or with neuraminidase resulted in loss of the inhibitory activity. Oligosaccharides released from Band 3 by hydrazinolysis effectively inhibited the binding of autologous IgG and 125I-labeled purified anti-Band 3 to the modified cells, whereas the oligosaccharides depleted of acidic components did not. Endo-beta-galactosidase and neuraminidase destroyed the activity of the oligosaccharides, but alpha-L-fucosidase did not. Furthermore, human lactoferrin that contains sialylated two N-acetyllactosaminyl units also exhibited potent inhibitory activity, and the activity was destroyed by endo-beta-galactosidase and neuraminidase. These results indicate that the antigenic determinants of Band 3 are located in sialylated poly-N-acetyllactosaminyl sugar chains. Based on this finding, mechanism of appearance of the antigen on senescent erythrocytes recognized by anti-Band 3 (senescent antigen) was discussed. 相似文献
16.
对我国11个地方鸡种1087羽鸡群进行了血型和血浆蛋白质多态7个位点22个等位基因的群体遗传学分析。结果表明,血型A、C位点,基因频率各鸡种存在显著差异,B位点则没有显著差异,血型和血浆蛋白质多态位点基因纯合系数普遍较低,血型因子分布在群体中较为分散,表明这些地方品种存在较大的选择潜力。血浆蛋白质多态位点,中国与外国的某些鸡种比较,存在着较大的相同或相异,可能意味着我国和外国鸡种起源上的同缘或品种起源、进化上的多样性。聚类分析表明,这11个地方鸡种可系统聚类成4个类群,其结果与目前人们对这些鸡种的表征分类不尽一致。 相似文献
17.
A blood-group Lewis precursor glycolipid was isolated from the plasma of a Lewis-negative individual [Le(a--b--)] and treated with fucosyltransferases from human gastric mucosa and GDP-fucose. Subsequently the glycolipid was adsorbed onto Le(a--b--) erythrocytes and the presence of blood-group Lewis antigens was assessed by passive hemagglutination with anti-Lewis sera. It was shown that the precursor glycolipid was enzymatically transformed to blood-group Lewis a (Lea) and Lewis b (Leb) specific glycolipids. Leb-glycolipid was also synthesized by fucosylation of an isolated Lea-glycolipid. Moreover Le(a--b--) erythrocytes were shown to develop Lea and Leb activities when subjected to enzymatic fucosylation, thus showing that Lewis-negative cells carry blood-group Lewis precursor glycolipid on the surface of their membrane. Le(a + b--) erthrocytes, upon enzymatic fucosylation, acquired Leb activity. 相似文献
18.
Eisuke P. Murono Amie L. Washburn 《The Journal of steroid biochemistry and molecular biology》1990,37(5):675-680
The present studies examined responses to hCG and/or insulin of 3 beta-hydroxy-5-ene-steroid dehydrogenase and steroid 5----4-ene-isomerase activity (3 beta-HSD) in cultured Band 2 and Band 3 cells from 25- to 40-day-old rats isolated on Percoll gradients. In Band 2 cells, from 25-day-old rats enzyme activity increased about 3- and 2.5-fold, after 6 days of exposure to hCG or insulin, respectively. However, hCG did not stimulate enzyme activity in Band 2 cells from 30-, 35- and 40-day-old animals, and responses to insulin alone or insulin plus hCG declined with age. In Band 3 cells only insulin increased enzyme activity at each age. Neither hCG or insulin altered DNA levels in Band 2 or Band 3 cells, suggesting that increased activity in Band 2 cells from 25-day-old rats was not due to cellular replication. However, hCG increased the number of cells staining positive for 3 beta-HSD about 4-fold in Band 2 cells from 25-day-old rats. Insulin did not increase the number of positive staining cells in Band 2 and Band 3 cells from 25-day-old rats, suggesting that its major effect was to increase enzyme activity in existing cells. These results suggest that during a limited period of maturation precursor cells in Band 2, which are undetected by histochemical staining for 3 beta-HSD, can be converted to Leydig cells in culture by hCG. 相似文献
19.
Na2SeO3 could affect the anion flux of Band 3 of inside-out erythrocyte membrane vesicles (IOVs). Such effect was believed to be
based on the interaction of SH groups of Band 3 with Na2SeO3. This effect could be eliminated when the cytoplasmic domain of Band 3 was proteolytically removed by trypsin. This suggested
that SH groups in the cytoplasmic domain were involved in such interaction. Measurement of the pH dependence of intrinsic
fluorescence intensity provided evidence that conformational changes of Band 3 occurred as a consequence of interaction with
selenite. KI quenching of intrinsic fluorescence of Band 3 could also show that there was a conformational change in the cytoplasmic
domain of Band 3 after reaction with Na2SeO3. Such conformational change in turn could be transmitted to the membrane domain of Band 3 monitored by quenching of intrinsic
fluorescence of Band 3 using hypocrellin B (HB) (a photosensitive pigment obtained from a parasitic fungus growing in Yunnan,
China).
It is suggested that the cytoplasmic domain of Band 3 is not necessary for its anion flux, but is essential for the regulation
(e.g., by Se) of its active site located at the membrane domain, and hence, it may provide evidence of communication between
the cytoplasmic domain and the membrane domain of Band 3. 相似文献
20.
用Triton X-100选择性抽提法,部分提纯了人红细胞膜带3蛋白。在除去去污剂后,带3蛋白被插入由磷脂酰胆碱/胆固醇组成的脂质体中。研究表明,用0.05%Triton抽提膜能除去大部分唾液酸糖蛋白,不会损失太多的带3蛋白。再用0.5%Triton抽提,可增溶出大量带3蛋白和少量其它膜蛋白。把它重组入脂质体后,在最后超离心步骤中得到了小的单层囊泡和大的多层囊泡。本文研究了单层重组囊泡输入~(35)SO_4~(2-)的功能,并和那些由人红细胞“发芽”得到的天然囊泡输入~(35)SO_肋~(2-)的功能作了比较。 相似文献