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1.
Antigenic Difference between Informofers and Protein bound to Polyribosomal mRNA from Rat Liver 总被引:4,自引:0,他引:4
NUCLEAR RNA with DNA-like base composition (heterogeneous nuclear RNA, Hn-RNA) is complexed with globular protein particles called informofers1–4. When mRNA is liberated from polysomes by EDTA or incubation with puromycin, it is isolated as a ribonucleoprotein complex (mRNP)5–8. Polyacrylamide gel electrophoresis has been used to study whether the informofer protein and the protein of polysomal mRNP are completely or partly identical9–11. The protein bound to haemoglobin mRNA is different from the informofer protein, but in rat liver and sheep thyroid polysomes a protein was found with characteristics similar to the informofer protein6,12,13. We have used a new immunological procedure to show that the proteins of the two ribonucleoprotein complexes in the rat liver are immunologically different. 相似文献
2.
THE first component (C1) in the complement system may be defined functionally as a macromolecule capable of binding to antigen-antibody complexes and inducing the sequential reactions of this system. C1 consists of three distinct proteins named C1q, C1r and C1s which,in serum, form a macromolecular complex held together by calcium ions1. The C1q protein was first isolated by Müller-Eberhard and Kunkel2 and Taranta et al.3. The ultrastructure of this basic, heat-labile 11S protein is outlined here. 相似文献
3.
Cloning of the sacS gene encoding a positive regulator of the sucrose regulon in Bacillus subtilis 总被引:1,自引:0,他引:1
Michel Débarbouillé Frank Kunst ré Klier Georges Rapoport 《FEMS microbiology letters》1987,41(2):137-140
The sacS gene controls the expression of 2 saccharolytic enzymes in Bacillus subtilis (sucrase and levansucrase).This paper describes a recombinant plasmid containing a mutant allele, sacSc. The plasmid was isolated from a B. subtilis DNA bank established in Escherichia coli. Moreover, it was shown that the sacSc allele, placed on a high-copy plasmid, is dominant over the wild-type chromosomal sacS+ allele. This result strongly suggests that the sacS gene encodes a positive regulatory protein. 相似文献
4.
Mice ranging in age from 16 to 44 days were injected intracerebrally with 3H-leucine, and incorporation into total brain proteolipids and the myelin proteolipid protein was measured. All proteolipids were isolated from whole brain by ether precipitation and separated into their individual components by SDS polyacrylamide gel electrophoresis. Two major proteolipids with apparent molecular weights of 20,700 and 25,400 were observed in these preparations, and their proportion increased over the developmental period examined. A Ferguson plot analysis comparing these proteins with those of isolated myelin showed that the 25,400-dalton proteolipid component from whole brain was the myelin proteolipid protein. Rates of incorporation of 3H-leucine into total brain proteolipids peaked at 22 days of age. Synthesis of the myelin proteolipid protein increased rapidly to a maximum value at 22 days and decreased rather slowly until at 44 days it was about 83% of its maximum rate of synthesis. The data indicate that the developmental pattern of synthesis of the myelin proteolipid protein is unlike that of the myelin basic proteins. Synthesis of the major myelin proteins is developmentally asynchronous in that peak synthesis of the myelin proteolipid appears to occur several days later than the basic proteins. In addition, it maintains its maximum rate of synthesis over a longer period of time than do the basic proteins. 相似文献
5.
Human myelin basic protein isolated from the brains of individuals who died with multiple sclerosis was more potent in inducing the aggregation of egg phosphatidylcholine vesicles than was the basic protein isolated from the brains of normal individuals. The portion of myelin basic protein which bound to egg phosphatidylcholine vesicles was separated from the free protein by sucrose density gradient centrifugation. Similar amounts of basic protein from normal or from multiple sclerosis brains are bound to the lipid and no consistent differences in the NG, NG dimethyl-arginine content of the protein fractions have been found. 相似文献
6.
The localization of proteins in myelin was studied by the use of a non-penetrating penetrating reagent. Tritiated 4,4′-diisothiocyano-2,2′-ditritiostilbene disulfonic acid was used to label the isolated myelin membrane. The membrane was labelled, the basic protein and the hydrophobic protein, lipophilin, were isolated. After 10 min of exposure to the reagent, the specific activity of lipophilin was found to be 10 times greater than that of the basic protein. Water shock did not alter the specific activities. However, sonication increased the specific activity of lipophilin but not that of basic protein. When the isolated proteins were labelled with 3H-labelled, 4,4′-diisothiocyano-2,2′-ditritiostilbene disulfonic acid, the specific activity of the basic protein was 10 times that of lipophilin. We concluded that the low specific activity of basic protein isolated from the labelled membrane was due to the inaccessible position of this protein in the membrane bilayer. 相似文献
7.
Kou Kubota 《Biochemical and biophysical research communications》1982,105(2):688-697
A growing organism that produces antibiotic peptide was incubated with L-(U-14C)serine for labeling linear gramicidin. Linear gramicidin was isolated by a simple chromatographic method from tyrothricin (mixture of linear gramicidin and tyrocidine) applied to a column of basic aluminum oxide. The hydrolysate of labeled linear gramicidin on thin layer chromatography showed that L-(U-14C)serine was one of a precursor of ethanolamine moiety by autoradiography. L-(3-14C)serine generated formic acid in the presence of tetrahydrofolic acid by an enzyme fraction prepared with ammonium sulfate, and further formed ethanolamine binding to the protein. Formylvaline was biosynthesized by it with tetrahydrofolic acid and ATP, and subsequently released from the protein. 相似文献
8.
9.
Molecular characterization of PeSOS1: the putative Na+/H+ antiporter of Populus euphratica 总被引:1,自引:0,他引:1
Populus euphratica is a salt-tolerant tree species growing in semi-arid saline areas. A Na+/H+ antiporter gene was successfully isolated from this species through RACE cloning, and named PeSOS1. The isolated cDNA was 3665 bp long and contained a 3438 bp open reading frame that was predicted to encode a 127-kDa protein
with 12 hypothetical transmembrane domains in the N-terminal part and a long hydrophilic cytoplasmic tail in the C-terminal part. The amino acid sequence of this PeSOS1 gene showed 64% identity with the previously isolated SOS1 gene from the glycophyte Arabidopsis thaliana. The level of protein expressed by PeSOS1 in the leaves of P. euphratica was significantly up-regulated in the presence of high (200 mM) concentrations of NaCl, while the mRNA level in the leaves
remained relatively constant. Immunoanalysis suggested that the protein encoded by PeSOS1 is localized in the plasma membrane. Expression of PeSOS1 partially suppressed the salt sensitive phenotypes of the EP432 bacterial strain, which lacks the activity of the two Na+/H+ antiporters EcNhaA and EcNhaB. These results suggest that PeSOS1 may play an essential role in the salt tolerance of P. euphratica and may be useful for improving salt tolerance in other tree species.
Yuxia Wu and Nan Ding contributed equally to this work. 相似文献
10.
Presence of Ornithine in the Urate-binding α<Subscript>1</Subscript>—<Subscript>2</Subscript>Globulin 总被引:1,自引:0,他引:1
THE urate-binding α1–α2 globulin has been isolated from human plasma in a highly purified state1. The protein was purified by DEAE-‘Sephadex’, ammonium sulphate precipitation and semi-preparative Polyacrylamide gel electrophoresis. The urate-binding α1–α2 globulin is a rod-shaped glycoprotein, containing 12.1% carbohydrate, with an isoelectric point of 4.6 and a molecular weight of 67,000 ± 4,000. Amino-acid analysis indicated an unknown basic compound which appeared as an extra peak just in front of lysine1. To identify this compound, high voltage paper electrophoresis has been carried out on a plate electrophoresis apparatus in pyridine-acetate buffer pH 3.5. A spot separated out corresponding to ornithine. Amino-acid analysis on a BC-200 automatic analyser (Bio-Cal Instruments Co., West Germany), with a 54 cm column at 55° C and with 0.35 M sodium citrate buffer, pH 5.28, as elution buffer at a flow-rate of 150 ml./h, showed that ornithine was present. The presence of ornithine in the protein hydrolysate was also verified by gas chromatography/mass spectrometry2. 相似文献
11.
Isolation and Characterization of a Protein from Sciatic Nerve Myelin responsible for Experimental Allergic Neuritis 总被引:5,自引:0,他引:5
EXPERIMENTAL allergic encephalomyelitis (EAE) is an autoimmune demyelinating disease1,2 of the central nervous system (CNS) induced by the basic Al protein3 of the myelin membrane (30% of the total protein). The complete aminoacid sequences of the human and bovine A1 proteins have been determined4,5. The open conformation6–8 of the A1 protein (18,400 molecular weight) emphasizes the role of the primary structure in determining its immunological properties. From isolated peptide fragments, disease-inducing sites of the A1 protein have been identified and subsequently synthesized9–11. 相似文献
12.
The covalent binding of metabolically activated 1,2-dibromoethane (DBE), a potent carcinogen, to chromatin constituents of forestomach and liver was examined . Chromatin was prepared from forestomach and liver of B6C3F1 mice and characterized. In order to activate DBE, microsomes and cytosol were isolated from mouse forestomach and liver and incubated with [14C]-DBE in the presence of a NADPH regenerating system. Results demonstrate that DBE bound covalently to the same extent to protein of microsomes and chromatin isolated from forestomach and liver. On the contrary, DBE bound significantly more to chromatin DNA of forestomach or liver than it did to salmon sperm DNA. It appears from these results that the metabolically activated DBE is more reactive to homologous DNA than exogenous DNA. Fractionation of DBE-bound chromatin protein into histone and nonhistone proteins resulted in higher binding of DBE to non-histone than to histone proteins isolated from forestomach and liver. 相似文献
13.
Comparison of Amino-acid Sequences of Hypothalamic Peptide,Brain-specific Histone and Myelin Basic Protein 总被引:2,自引:0,他引:2
SEVERAL studies have described the basic proteins of porcine central nervous tissue. Shome and Saffran1 have isolated and purified a large peptide (or small protein) of molecular weight about 14,000 from acetone-dried hog hypothalamic powder. Sixteen of the approximately twenty-six tryptic digest fragments of this protein have been sequenced. The protein has no pressor or ACTH-releasing activity. Tomasi-and Kornguth2 purified and partially characterized a basic protein from pig brain and, on the basis of fluorescent antibody studies, they concluded that this protein is a brain-specific histone, found in neurone nuclei (or nucleoli) of several animals3–5. 相似文献
14.
《Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology》1993,1161(2-3):187-193
The peptide fragment Pro212-Ile276 of human protein C was produced as a part of a fusion protein in Escherichia coli. The identity of the peptide was confirmed by immunoblotting experiments using specific antibodies to intact protein C. The peptide Pro212-Ile276 was isolated from the fusion protein after mild hydrolysis with formic acid by gel filtration and reverse-phase HPLC. This peptide fragment was used to produce antibodies specific for the heavy chain of protein C which recognized native protein C present in blood plasma. Antibodies to intact protein C reacted also with the Pro212-Ile276 peptide fragment, indicating that this region is immunogenic in intact protein C and may represent a native epitope. 相似文献
15.
Isolation and characterization of a Na+/H+ antiporter gene from the halophyte Atriplex gmelini 总被引:25,自引:0,他引:25
Hamada Akira Shono Mariko Xia Tao Ohta Masaru Hayashi Yasuyuki Tanaka Akira Hayakawa Takahiko 《Plant molecular biology》2001,46(1):35-42
With a homologous gene region we successfully isolated a Na+/H+ antiporter gene from a halophytic plant, Atriplex gmelini, and named it AgNHX1. The isolated cDNA is 2607 bp in length and contains one open reading frame, which comprises 555 amino acid residues with a predicted molecular mass of 61.9 kDa. The amino acid sequence of the AgNHX1 gene showed more than 75% identity with those of the previously isolated NHX1 genes from glycophytes, Arabidopsis thaliana and Oryza sativa. The migration pattern of AgNHX1 was shown to correlate with H+-pyrophosphatase and not with P-type H+-ATPase, suggesting the localization of AgNHX1 in a vacuolar membrane. Induction of the AgNHX1 gene was observed by salt stress at both mRNA and protein levels. The expression of the AgNHX1 gene in the yeast mutant, which lacks the vacuolar-type Na+/H+ antiporter gene (NHX1) and has poor viability under the high-salt conditions, showed partial complementation of the NHX1 functions. These results suggest the important role of the AgNHX1 products for salt tolerance. 相似文献
16.
A protein kinase specific for casein and acidic ribosomal proteins was isolated and partly characterized.It was found that the enzyme utilizes GTP and ATP as phosphoryl donors. Its affinity for ATP was considerably higher than for GTP with the km values of 7.6 × 10-6M and 5.5 × 10-5M, respectively.Two-dimensional acrylamide gel electrophoresis revealed the phosphorylation of the same ribosomal proteins with either of the [-32P] nucleotides used. It was also shown that one acidic protein (S1 or S2) of 40 S and two acidic proteins (L2 and L3) of 60 S ribosomal subunits were predominantly phosphorylated in vitro. The phosphorylated proteins: L2 and L3 seem to correspond to the proteins of L7 and L12 of E. coli ribosomes. The isolated kinase phosphorylated several basic ribosomal proteins though to a lower extent than the acidic ones. 相似文献
17.
By reducing the amount of ginkgo water-soluble polysaccharides, which occupy about 35% of the wet seed mass and interfere with the extraction of RNA, cDNA-quality mRNA was obtained from developing seeds of Ginkgo biloba. Based on the NH2-terminal 17-amino acid sequence and an internal 12-amino acid sequence derived from the basic subunit of ginnacin, 11S-seed storage protein family of ginkgo, two degenerate oligonucleotide primers were synthesized and used for polymerase chain reaction (PCR). The resulting PCR product was used for screening the above endosperm cDNA library, and a plaque carrying the 1614 bp cDNA insert, which contained the entire coding region for a precursor of ginnacin was isolated. This is the first reported cloning of cDNA from ginkgo seeds. The deduced primary sequence is composed of a signal peptide segment (25 amino acid residues) and an acidic subunit (248 residues) followed by a basic subunit (187 residues). It was also found that the post-translational cleavage site in the ginnacin precursor is the Asn-Asn rather than the Asn-Gly bond found in a variety of the major subunit precursors in 11S seed protein family known to date. We showed that a purified soybean extract and an extract of ginkgo seeds can specifically hydrolyze-Asn248-Asn249- but not -Asn249-Val250-, in the heptapeptide Gly-Asn248-Asn-Val-Glu-Glu-Leu that corresponds to the ginnacin cleavage region.Abbreviations SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- PVDF
polyvinylidene difluoride
- CBB
Coomassie Brilliant Blue
- HPLC
high-performance liquid chromatography
- bp
base pair(s)
- PCR
polymerase chain reaction 相似文献
18.
The partial removal of tightly bound Ca2+ from dialysed neem (Azadirachta indica) gum, resulted in the release of a basic protein from a highly anionic polysaccharide-protein complex as evidenced by chromatographic
studies on TEAE-cellulose. Complete removal of Ca2+ caused, in addition, the release of a minor heteropolysaccharide which was found in association with the basic protein. These
processes were reversed on the addition of Ca2+. The gum, in addition, contained a protein-rich component accounting for 35% protein and 7.5% total carbohydrate. This component
behaved as a distinct entity during ion-exchange chromatography of the native gum solutions, or which were either partially
or completely depleted of bound Ca2+. 相似文献
19.
Lopes-Ferreira M Magalhães GS Fernandez JH Junqueira-de-Azevedo Ide L Le Ho P Lima C Valente RH Moura-da-Silva AM 《Biochimie》2011,93(6):971-980
Lectins are glycan-binding receptors that recognize glycan epitopes on foreign pathogens and in the host systems. They can be involved in functions that include innate immunity, development, immune regulation and homeostasis. Several lectins have been purified and characterized from fish species. In this work, using cation-exchange chromatography, a galactose-specific lectin belonging to the family of C-type lectins was isolated from the venom of the Brazilian venomous fish Thalassophryne nattereri. Nattectin is a basic, non-glycosilated, 15 kDa monomeric protein. It exhibits hemagglutination activity that is independent of Ca2+. We also demonstrated a lectin activity for Nattectin in the innate immune system, especially in neutrophil mobilization in mice, indicating that marine organisms are source of immunomodulator agents. 相似文献
20.
Extracellular Black Yeast Polysaccharide composed of N-Acetyl Glucosamine and N-Acetyl Glucosaminuronic Acid 总被引:2,自引:0,他引:2
ALLENE JEANES KERMIT A. BURTON MARTIN C. CADMUS CLARENCE A. KNUTSON GERALD L. ROWIN PAUL A. SANDFORD 《Nature: New biology》1971,233(43):259-260
POLYSACCHARIDES containing N-acylated hexosaminuronic acids as major components have been isolated from bacterial cells1–4,7 and capsules5,6. Most of these polysaccharides are antigens from pathogens1,2,4,5,7. Polysaccharides from cells or pathogens, however, are unsuitable for investigations of the basic properties and possible practical value of biopolymers, because they are recovered with difficulty and in low yields. It is significant, therefore, that we have isolated in good yield from cultures of an unidentified black yeast (apparently a non-pathogen) an extracellular polysaccharide composed of residues of N-acetyl glucosamine (GlcNAc) and N-acetyl glucosaminuronic acid (GlcNAcUA) in a molar ratio of approximately 2:1. The composition and extracellular occurrence of this polysaccharide make it unique among those from microorganisms. It is the first practicably obtainable member of a new class of biopolymers to be characterized1–7. Additional members of this class are foreseen from our exploratory research. 相似文献