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1.
A new type of dihydropteridine reductase [EC 1.6.99.10], which is specific for NADPH as the substrate in the reduction of quinonoid-dihydropterin to tetrahydropterin, was purified to homogeneity from bovine liver and human liver. The molecular weight of the enzyme was determined to be 65,000-70,000. The enzyme was composed of two subunits with identical molecular weight of 35,000; the amino terminal residue was determined to be valine. The isoelectric point of the enzyme was 7.05. The physicochemical properties of this enzyme were quite different from those of bovine liver NADH-specific dihydropteridine reductase [EC 1.6.99.7]. NADPH-specific dihydropteridine reductase did not cross-react with an antiserum raised against the NADH-specific dihydropteridine reductase, nor did the latter enzyme react with an antiserum to the former enzyme, indicating that the two enzymes have no common antigenic determinants. NADPH-specific dihydropteridine reductase from human liver was shown to have properties similar to those of the bovine liver enzyme. 相似文献
2.
Comparison of cytosol retinol binding proteins from bovine retina, dog liver, and rat liver 总被引:1,自引:0,他引:1
Cytosol retinol binding proteins (CRBP's) have been purified from rat liver, dog liver, and bovine retina. All had identical molecular weights on sodium dodecyl sulfate electrophoresis. They had different RF values on non-sodium dodecyl sulfate gels at pH 8.9. The three CRBP exhibited similar absorption and fluorescence spectra. The absorbance of the ligand was perturbed after binding, the main band shifting bathochromically and exhibiting a lambda(max) at 350 nm compared with 328 nm for free retinol in hexane. Additionally, subsidiary peaks appeared at 335 and 367 nm. Rabbit antiserum against rat liver CRBP cross-reacted with CRBP's from dog liver and bovine retina. The Ouchterlony immunodiffusion technique indicated that these proteins have molecular structures with identical antigenic determinants. All three CRBP's had amino acid composition that were virtually identical, as judged by our own observations and those of other laboratories. The molecular structure of cytosol retinol binding proteins appears to be highly conserved, irrespective of species or tissue of origin. 相似文献
3.
The present experiments showed that the guinea pig antiserum prepared against the main polypeptides of 14 S dynein from Tetrahymena cilia reacted with sea urchin sperm flagellar dynein and with bovine brain high molecular weight protein to give rise to a precipitin line confluent with that formed between the antiserum and Tetrahymena dynein. Furthermore, it was found that this antiserum also reacted with tubulins from Tetrahymena cilia, sea urchin sperm flagella and bovine brain to give rise to the confluent precipitin line. Among muscle proteins, only actin preparation from rabbit skeletal muscle reacted with the anti-Tetrahymena dynein serum, whereas neither rabbit skeletal muscle myosin, chicken skeletal muscle tropomyosin nor chicken skeletal muscle troponin reacted with the antiserum. These results suggest that dynein and tubulin and probably actin share an antigenic determinant regardless of different protein species and of different animal species. The common antigenic determinant was detected only when the proteins denatured with urea/sodium dodecyl sulfate/beta-mercaptoethanol/N-ethylmaleimide were used, but it was not detected at all when the native proteins were used. This implies that a certain common antigenic determinant which is involved in the precipitin line formation exists in the primary structures of dyneins and tubulins and probably actin, and is hidden inside the tertiary structures of the native protein molecules. 相似文献
4.
L A Lokshina I A Tarkhanova O N Lubkova N V Golubeva T A Gureeva 《Biulleten' eksperimental'no? biologii i meditsiny》1983,96(10):50-53
Rabbit antisera were prepared against three highly purified enzymes from bovine spleen: proteinase I (cathepsin L), proteinase II (cathepsin H), and cathepsin B. The Ouchterlony double diffusion test shows that each antiserum specifically reacts with the corresponding antigen and does not cross react with other proteinases. These data provide evidence that the three proteinases are distinct with respect to their antigenic properties. Using specific antisera, the identity of two preparations of proteinase I isolated by different methods was demonstrated. Analysis of the fractions obtained in the course of isolation procedure revealed a component reacting with antisera against proteinase I. It had a greater molecular mass than proteinase I (30 000-40 000), was richer in antigenic respect and had a lower proteolytic activity as compared with proteinase I. The effect of various inhibitors and denaturation conditions on antigenic properties of proteinases was also studied. 相似文献
5.
An antiserum to rat liver catechol-O-methyltransferase (COMT) was utilized in the immunological characterization of COMT from rat kidney, brain, and choroid plexuses, in addition to rat liver. The presence of anti-COMT activity was confirmed by the direct inhibition of the activity of the enzyme from rat liver by small quantities of the antiserum and by the inhibition of the activity of the enzyme from rat brain. The specificity of the antiserum was demonstrated both by immunoelectrophoresis of rat liver COMT, and by a partial purification of rat liver COMT in which changes in COMT specific activity were correlated with the appearance of a precipitin line in double-immunodiffusion experiments. The antigenic similarity of the enzyme derived from rat liver, kidney, brain, and choroid plexuses was demonstrated by the formation of a precipitin line of identity when preparations from these four tissues were diffused against the antiserum. 相似文献
6.
J. M. Grondona P. Fernández-Llebrez J. Pérez M. Cifuentes J. M. Pérez-Fígares E. M. Rodríguez 《Cell and tissue research》1994,276(3):515-522
We have raised antisera against extracts of the subcommissural organ (SCO) of the dogfish, Scyliorhinus canicula L. Brains of 2900 specimens were collected in acetone, and the region containing the SCO and posterior commissure was removed and extracted in three different media. Antisera against these crude extracts were raised in rats and rabbits. Sequential absorptions of the antisera with extracts from different regions of the dogfish brain were performed to eliminate unwanted antibodies. When used to immunostain sections of the whole central nervous system of the dogfish, these purified antisera reacted selectively with the SCO-Reissner's fiber complex. An antiserum against bovine Reissner's fiber was also used. The antisera against the dogfish SCO and bovine Reissner's fiber showed the same staining pattern in the SCO and the Reissner's fiber of the dogfish. For comparative purposes, the brains of 15 vertebrate species from all vertebrate classes were immunostained with both antisera. The anti-dogfish SCO serum reacted with the SCO of the dogfish and that of other phylogenetically related elasmobranch species. Neither the SCO of a primitive elasmobranch species, Heptranchias perlo, nor the SCO of the other classes of vertebrates reacted with the anti-dogfish SCO serum. However, the antiserum against bovine Reissner's fiber reacted with the SCO of all the investigated species. It is concluded that some epitopes (or compounds) in the secretory material of the SCO are class-specific, whereas others are conserved and are synthesized by the SCO in most vertebrate species. 相似文献
7.
M Cherkaoui Malki A Kante C Demigne N Latruffe 《Comparative biochemistry and physiology. B, Comparative biochemistry》1992,101(3):413-420
1. The properties of rat liver and bovine heart R-3-hydroxybutyrate dehydrogenase (BDH) have been extensively studied in the past 20 years, but little is known concerning the biogenesis and the regulation of this dehydrogenase over different species. 2. In addition, controversial results were often reported concerning the activity, the level and the subcellular location of this enzyme in ruminants. 3. BDH activity found in liver and kidney mitochondria from ruminants (cow and sheep) is low, while it is much higher in rat. 4. However, the enzyme activity is detected in microsomes and in cytosol of liver and of kidney cells from ruminants. These activities are not correlated to ketonaemia level. 5. Although low BDH activity is detected in liver mitochondria from ruminants; the bovine liver BDH gene seems to be translated since BDH can be immunodetected by using an antiserum raised against bovine heart BDH. 6. Beside this, the good cross-reactivity between heart BDH and liver BDH suggests their high level of homology in ruminants. 相似文献
8.
Tissue-specificity overrides species-specificity in cytoplasmic cytochrome c oxidase polypeptides 总被引:4,自引:0,他引:4
With a high-resolving dodecyl sulfate electrophoretic system rat liver cytochrome c oxidase was separated into 13 different polypeptides. An antiserum against rat liver holocytochrome c oxidase immunoreacted with all 13 polypeptides, as demonstrated by immunofluorescence after transfer of the separated Coomassie blue-stained bands on nitrocellulose and coupling with FITC-protein A ("western blot"). Polypeptide-specific antisera reacted only with their corresponding polypeptides indicating that the various protein bands are represented by individual polypeptides. From total proteins of rat liver, kidney, heart, spleen and skeletal muscle mitochondria, only the cytochrome c oxidase polypeptides showed immunofluorescence with an antiserum against the rat liver holoenzyme. In contrast to the polypeptide from liver, polypeptide VIa from heart and skeletal muscle showed little or no reactivity, indicating a tissue-specificity of this polypeptide. Mitochondrial proteins from pig, bovine and blackbird heart were incubated with an antiserum against the rat liver holoenzyme. Immunoreaction was found with most cytochrome c oxidase polypeptides but not with polypeptide VIa. This result demonstrates less immunological relationship between tissue-specific polypeptides (VIa, VIIa and VIII) of the same species than between tissue-unspecific polypeptides of different species. 相似文献
9.
Investigations carried out with the use of a CELIA system have revealed that antibodies in the sera of patients with primary erysipelas and antibodies in rabbit antiserum to the ribosomes of group A streptococcus specifically bind with adsorbed streptococcal ribosomes, recognizing the antigenic determinants of streptococcal ribosomes, which differ from those of individual Gram-negative prokaryotes (Escherichia coli, Shigella sonnei, Shigella flexneri, Salmonella minnesota). The modified CELIA system used in this investigation has made it possible to find out that antibodies in the sera of patients with primary erysipelas and antibodies in rabbit ribosomal antiserum bind with different antigenic determinants of the ribosomes of group A streptococcus. 相似文献
10.
Molecular and immunological comparison of human dihydropteridine reductase in liver, cultured fibroblasts and continuous lymphoid cells. 总被引:2,自引:0,他引:2
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An antiserum was raised in a rabbit against highly purified human liver dihydropteridine reductase (EC 1.6.99.7). Dihydropteridine reductase from human liver, in human cultured fibroblasts and in continuous lymphoid cells all showed identical antigenic properties. The structural characteristics of the reductase from these three sources were further compared by the use of high-precision two-dimensional polyacrylamide-gel electrophoresis. The enzyme from radiolabelled fibroblasts and continuous lymphoid cells was isolated by immunoprecipitation or by affinity chromatography and compared with the purified liver enzyme. Two major polypeptide species were resolved, and polypeptides from all three sources co-migrated identically. Indirect evidence is presented indicating that one of the polypeptide species may have been derived from the other via a post-translational modification. These results support the concept that the same structural gene(s) encodes for dihydropteridine reductase in human liver, fibroblasts and lymphocytes. 相似文献
11.
The bovine mitochondrial system is being developed as a model system for studies on mammalian mitochondrial ribosomes. Information is emerging on the structural organization and RNA binding properties of proteins in these mitochondrial ribosomes. Unexpectedly, these ribosomes appear to interact directly with GTP, via a high affinity binding site on the small subunit. Despite major differences in their RNA content and physical properties, mammalian mitochondrial and cytoplasmic ribosomes contain about the same number of proteins. The proteins in each kind of ribosome have a similar size distribution, and both sets are entirely coded by nuclear genes, raising the possibility that these different ribosomes may contain the same set of proteins. Comparison of bovine mitochondrial and cytoplasmic r-proteins by co-electrophoresis in two-dimensional gels reveals that most of the cytoplasmic ribosomal proteins are more basic than the mitochondrial ribosomal proteins, and that none are co-migratory with mitochondrial ribosomal proteins, suggesting that the proteins in the two ribosomes are different. To exclude the possibility that the electrophoretic differences result only from post-translational modification of otherwise identical proteins, antibodies against several proteins from the large subunit of bovine mitochondrial ribosomes were tested against cytoplasmic ribosomes by solid phase radioimmunoassay and against cytoplasmic ribosomal proteins on Western blots. The lack of cross-reaction of these antibodies with cytoplasmic r-proteins suggests that mitochondrial ribosomal proteins have different primary structures and thus are most likely encoded by a separate set of nuclear genes. 相似文献
12.
The existence of different antigenic determinants in the beef heart adenine nucleotide (AdN) carrier was demonstrated by exploring the reactivity of fragments of the carrier protein with rabbit antisera directed against either the beef heart AdN carrier denatured with sodium dodecyl sulfate (NaDodSO4 carrier) or the beef AdN carrier liganded by the specific inhibitor carboxyatractyloside (CATR-carrier). The antigenic determinants reacting with antiserum to the CATR-carrier appeared to be close to the N- and C-terminal ends of the carrier protein, whereas those reacting with antiserum to the NaDodSO4 carrier were located preferentially in the central region of the protein. The same antisera were used to study the immunogenic specificity of the beef liver AdN carrier, the rat heart AdN carrier, and the rat liver AdN carrier. The beef liver and rat heart AdN carriers were found to react with both antisera whereas the rat liver AdN carrier reacted essentially with the CATR-carrier antiserum. 相似文献
13.
Cardiac contractile protein phosphatases. Purification of two enzyme forms and their characterization with subunit-specific antibodies 总被引:13,自引:0,他引:13
M C Mumby K L Russell L J Garrard D D Green 《The Journal of biological chemistry》1987,262(13):6257-6265
Two forms of protein phosphatase which dephosphorylate cardiac myosin or myosin light chains and the inhibitory subunit of cardiac troponin were purified from bovine cardiac muscle. The enzymes were composed of subunits of Mr = 63,000, 55,000, and 38,000 in a 1:1:1 molar ratio (PT-1) or Mr = 63,000 and 38,000 in a 1:1 molar ratio (PT-2). Native gel electrophoresis and sucrose gradient sedimentation indicated that activity toward all three substrates was due to a single enzyme species. A monoclonal antibody and polyclonal antiserum directed against an Mr = 38,000 protein phosphatase from this tissue specifically reacted with the Mr = 38,000 subunit of PT-1 and PT-2. The specificity of antibodies for the Mr = 38,000 subunit indicated that it was distinct from the other subunits. The Mr = 63,000 subunits of PT-1 and PT-2 were identical based on mobility on sodium dodecyl sulfate gels and one-dimensional peptide maps. Specificity of antiserum against the Mr = 55,000 subunit of PT-1 showed that this subunit was a distinct protein and not derived from the Mr = 63,000 subunit by proteolysis. PT-2 but not PT-1 could interact with antiserum against the Mr = 38,000 catalytic subunit in competitive immunoassays indicating that the presence of the Mr = 55,000 subunit may alter or mask antigenic site(s). Analysis of the enzymatic properties of PT-1 and PT-2 showed that PT-2 had higher activity with myosin, myosin light chains, and phosphorylase while PT-1 had higher activity with troponin. The results indicate that the presence of the Mr = 55,000 subunit may alter the enzymatic properties of the catalytic subunit. 相似文献
14.
Antibodies were raised in rabbits against 70S ribosomes, 50S and 30S ribosomal subunits individually. Purified immunoglobulins
from the antiserum against each of the above ribosomal entities were tested for their capabilities of precipitating 70S, 50S
and 30S ribosomes. The observations revealed the following: (i) The antiserum (IgG) raised against 70S ribosomes precipitates
70S ribosomes completely, while partial precipitation is seen with the subunits, the extent of precipitation being more with
the 50S subunits than with 30S subunits; addition of 50S subunits to the 30S subunits facilitates the precipitation of 30S
subunits by the antibody against 70S ribosomes. (ii) Antiserum against 50S subunits has the ability to immunoprecipitate both
50S and 70S ribosomes to an equal extent. (iii) Antiserum against 30S subunits also has the property of precipitating both
30S and 70S ribosomes. The differences in the structural organisation of the two subunits may account for the differences
in their immunoprecipitability. 相似文献
15.
Antibodies have been prepared against the 2-oxoglutarate transport proteins purified from bovine heart and rat liver mitochondria. The anti-heart antiserum cross-reacts with the 2-oxoglutarate carrier (OGC) from beef, pig, rat and rabbit heart, but not with the OGC from liver of the same animals. Conversely, the anti-liver antiserum recognizes the carrier protein from liver of all species tested but not from heart. Immunoinactivation of oxoglutarate transport activity by the antibodies is also tissue specific. Peptide maps of purified OGC show structural differences between the carrier from heart and liver of the same animal species. These results indicate the existence of isoforms of the OGC in heart and liver. 相似文献
16.
Takao Yokota Satoru Watanabe Yoichi Ogino Isomaro Yamaguchi Nobutaka Takahashi 《Journal of Plant Growth Regulation》1990,9(1-3):151-159
Antiserum against the brassinosteroid (BR), castasterone, was produced by immunizing a rabbit with castasterone-carboxymethoxylamine oxime conjugated with bovine serum albumin (BSA). In a radioimmunoassay (RIA), the antiserum recognized a range of naturally occurring BRs with varying specificities. Detection limits of castasterone and brassinolide were approximately 0.3 pmol. This RIA system was successfully used for analyzing endogenous BRs in seeds and stems ofPhaseolus vulgaris L., and showed that stems are quite different from seeds in terms of the species and quantity of the endogenous BRs. 相似文献
17.
Benoît Verjans Frederic Hollande Colette Moreau Claudine Lejeune Christophe Erneux 《Cellular signalling》1990,2(6):595-599
Inositol 1,4,5-trisphosphate 5-phosphatase catalyses the dephosphorylation of the phosphate in the 5-position from inositol 1,4,5-trisphosphate and inositol 1,3,4,5-tetrakisphosphate. One particulate and two soluble enzymes were previously described in bovine brain. In this study, we have obtained a precipitating antiserum against soluble type I inositol 1,4,5-trisphosphate 5-phosphatase. The particulate, but not the soluble type II enzyme, was immunoprecipitated by the serum. Inositol 1,4,5-triphosphate 5-phosphatase activity from crude extracts of rat brain, human platelets and rat liver were immmunoprecipitated by the same antibodies, suggesting the existence of common antigenic determinant among inositol 1,4,5-trisphosphate 5-phosphatases of diverse sources. 相似文献
18.
Immunological evidence for structural homology between Drosophila melanogaster (S14), rabbit liver (S12), Saccharomyces cerevisiae (S25), Bacillus subtilis (S6), and Escherichia coli (S6) ribosomal proteins. 总被引:2,自引:1,他引:1
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Specific antibodies directed against Drosophila melanogaster acidic ribosomal protein S14 were used in a comparative study of eucaryotic and procaryotic ribosomes by immunoblotting and enzyme-linked immunosorbent assays. Common antigenic determinants and, thus, structural homology were found between D. melanogaster, Saccharomyces cerevisiae (S25), rabbit liver (S12), Bacillus subtilis (S6), and Escherichia coli (S6) ribosomes. 相似文献
19.
P. H. Graham 《Archives of microbiology》1971,78(1):70-75
Summary Antisera were prepared against cell material from 3 strains of A. radiobacter and 6 of A. tumefaciens. Agar diffusion and immune absorption techniques revealed 3 antigens common to each strain of these organisms. However, 5 different lipopolysaccharide antigens occurred in the 9 test strains. There was no obvious species differences in the distribution of these antigens. Mannose, and possibly glucuronic acid were immunologically active in one of the lipopolysaccharides.The agrobacterial antisera were further crosstested with antigenic material for 34 strains of Rhizobium. Fast-growing rhizobia showed extensive cross-reaction, but only one of 7 R. lupini strains tested reacted with any antiserum. 相似文献
20.