Showing papers on "Gel electrophoresis published in 1969"
TL;DR: A high resolution gel electrophoresis of histone is described, capable of distinguishing between histone fractions whose mobilities differ by as little as 1% under the conditions of pH and urea concentration employed.
2,292 citations
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01 Jan 1969
TL;DR: In this paper, the authors performed electrophoresis of proteins in polyacrylamide and starch gels, and showed that proteins in these gels can be used for protein synthesis.
Abstract: Electrophoresis of proteins in polyacrylamide and starch gels , Electrophoresis of proteins in polyacrylamide and starch gels , مرکز فناوری اطلاعات و اطلاع رسانی کشاورزی
216 citations
TL;DR: Vinblastine quantitatively precipitates a protein from supernatants obtained from high-speed centrifugation of homogenates of HeLa cells and of pig brain that migrates as a single band on gel electrophoresis, has a mobility identical to that of purified microtubule protein, and-like micro Tubule protein-binds colchicine.
Abstract: Vinblastine quantitatively precipitates a protein from supernatants obtained from high-speed centrifugation of homogenates of HeLa cells and of pig brain. This protein migrates as a single band on gel electrophoresis, has a mobility identical to that of purified microtubule protein, and-like microtubule protein-binds colchicine. The precipitation is partially inhibited by 0.9 percent NaCl.
194 citations
TL;DR: Experiments with S35-labeled SDS suggest that the SDS-protein complex is a loose one and that exchange of SDS with the environment occurs at an exceedingly high rate.
190 citations
TL;DR: Treatment of the WSN strain of influenza virus with the non-ionic detergent nonidet P-40, followed by velocity gradient centrifugation, yielded a ribonucleoprotein (RNP) with a sedimentation constant of about 38 S which contained the five pieces of RNA previously found by extracting the virus with phenol-SDS.
186 citations
TL;DR: Preparations of bovine pancreatic deoxyribonuclease obtained by the ammonium sulfate precipitation procedure of Kunitz can be further purified by chromatography on sulfoethyl-Sephadex at pH 4.70 and show that the enzyme contains glucosamine and mannose and establish DNase as a glycoprotein.
165 citations
TL;DR: A temperature-sensitive mutant of E. coli, which cannot synthesize DNA at high temperature but can continue cell division, shows a difference in a membrane protein fraction at highTemperature by gel electrophoresis, which may link DNA replication and cell division through the membrane.
Abstract: A temperature-sensitive mutant of E. coli, which cannot synthesize DNA at high temperature but can continue cell division, shows a difference in a membrane protein fraction at high temperature by gel electrophoresis. This mutation of a bacterial membrane protein is unique. The protein may link DNA replication and cell division through the membrane.
163 citations
TL;DR: Chromatography of extracts of membrane proteins on Sephadex G-150 equilibrated with buffers containing sodium dodecyl sulfate was found to be a useful method of fractionation, which also leads to an effective separation of phospholipid from these proteins.
143 citations
TL;DR: Of six proteins identified in virions of the New Jersey serotype, only the smallest protein (P6) could be distinguished from any of the six proteins of the Indiana serotype on the basis of migration in SDS gels.
Abstract: Three major and three minor structural proteins were identified by polyacrylamide gel electrophoresis of purified infectious virions of the Indiana serotype of vesicular stomatitis (VS) virus disrupted with acetic acid, 0.5 m urea, sodium dodecyl sulfate (SDS), and 2-mercaptoethanol. Molecular weights of the six virion proteins were estimated by comparative electrophoretic migration of known marker proteins in the presence of SDS. The following values were obtained: major proteins P6 congruent with 34,500, P5 congruent with 59,500, and P4 congruent with 81,500; minor proteins P3 congruent with 140,000, P2 congruent with 186,000, and P1 congruent with 275,000. P1 did not disaggregate in 8 m urea, but P2 and P3 did. The possibility that P1 is an uncleaved large polypeptide chain could not be ruled out. Six identical protein components were dissociated from Indiana VS virions grown in chick and mouse cells; no cellular proteins could be detected in purified virions. Of six proteins identified in virions of the New Jersey serotype, only the smallest protein (P6) could be distinguished from any of the six proteins of the Indiana serotype on the basis of migration in SDS gels. The defective T particles of Indiana VS virus contained the same six proteins in essentially the same proportions as those of the infectious B virions. Only P6 and P5 could be cleanly separated by preparative gel electrophoresis.
109 citations
TL;DR: MYOSIN from rabbit skeletal muscle contains low molecular weight proteins (light chains) as well as the two large polypeptide chains which comprise the bulk of the molecule.
Abstract: MYOSIN from rabbit skeletal muscle contains low molecular weight proteins (light chains) as well as the two large polypeptide chains which comprise the bulk of the molecule. Alkali treatment has been used1–4 to separate the light from the heavy chains, and they have been characterized by gel electrophoresis and in the ultracentrifuge. The molecular weights are still in dispute, being reported as 20,000 daltons2 and 32,000 daltons3, but the light chains give at least three bands on gel electrophoresis and these molecular weight determinations2,3 were made with the mixture.
100 citations
TL;DR: It is suggested that V SP-3 is a glycoprotein and a major component of the viral envelope, that VSP-2 is the protein moiety of the nucleoprotein core, and that Vsp-1 may represent a capsid protein tightly bound to the envelope.
TL;DR: Electrophoresis experiments suggested preferential pronase attack on some higher molecular weight membrane proteins; these same electrophoretic components were preferentially released at very low concentrations of sodium dodecyl sulfate,, indicating differences in the organization of proteins within the membrane.
TL;DR: Gradient gels are studied in which protein molecules are driven through progressively decreasing pores until they are brought to a near dead stop in order of their size; a procedure appropriately called “pore-limit electrophoresis”5.
Abstract: ELECTROPHORETIC migration of proteins depends on their electrophoretic mobility and the resistance of the supporting medium. The latter is an important factor in starch and polyacrylamide gels the action of which as molecular sieves contributes greatly to the resolution of protein mixtures. This is further enhanced in gradient gels1–6 in which protein molecules are driven through progressively decreasing pores until they are brought to a near dead stop in order of their size; a procedure appropriately called “pore-limit electrophoresis”5.
TL;DR: The apparatus is manufactured by LKB-Produkter AB, Stockholm, Sweden, and the resolution is close to that obtained in analytical gel electrophoresis runs.
TL;DR: A reproducible procedure for the large scale purification of pig heart supernatant malate dehydrogenase which yields up to 400 mg of homogeneous protein has been developed and results indicate that pigheart supernatants malate dehydration is made up of two identical or very similar subunits.
TL;DR: A new catalyst system for polyacrylamide gels at low pH has been developed that is more effective and not inhibited by molecular oxygen.
TL;DR: In this degree of homology, the aldolases resemble the glyceraldehyde 3-phosphate dehydrogenases from various sources, suggesting that such homology may be a common feature of glycolytic enzymes.
Abstract: 1
The primary structures of fructose diphosphate aldolases from the skeletal muscle of rabbit, pig, ox and sturgeon have been compared by means of zone electrophoresis, amino acid and N-terminal analysis, peptide mapping and cyanogen bromide cleavage.
2
The S-carboxymethylated mammalian enzymes give two bands on polyacrylamide gel electrophoresis, under conditions where the S-carboxymethylated sturgeon enzyme gives only one. The two bands usually stain unequally with the rabbit enzyme, whereas with the ox and pig enzymes a more equal intensity is observed.
3
The mammalian enzymes are highly homologous by the criteria used, and it seems likely that the sturgeon enzyme will be fairly closely alike. In this degree of homology, the aldolases resemble the glyceraldehyde 3-phosphate dehydrogenases from various sources, suggesting that such homology may be a common feature of glycolytic enzymes.
4
Despite the occurrence of two bands in gel electrophoresis of the mammalian enzymes, it is clear that there cannot be substantial sequence differences between the subunit peptide chains of a given aldolase. Such differences, if they exist, must be relatively few in number. Other explanations, such as chemical modification of some of the chains, may account for the observed electrophoretic behaviour in the mammalian enzymes.
5
The ox enzyme contains one additonal cysteine residue per subunit compared with the other mammalian enzymes. This change has been located in the primary structure and since the cysteine residue is reactive in the native ox enzyme, it may provide a useful site for attachment of heavy atoms in X-ray crystallographic studies.
TL;DR: Ribonucleases may be detected in polyacrylamide gels after electrophoresis by incubation in a solution of RNA, followed by a 30 second dip into 0.2% toluidine blue, which is sensitive to small amounts of plant RNases.
TL;DR: The results indicate that the upper band of the Ficoll gradient consists mainly of mechanically disrupted cytoplasmic membrane.
TL;DR: Polyacrylamide gels containing 15% acrylamides provide a molecular sieve of unique resolving power for electrophoretic separation of low molecular weight RNA on the basis of size, and the exact relation between mobility and sedimentation rates is reported.
TL;DR: Polyacrylamide gel electrophoresis has been used to fractionate mixtures of oligomers produced by enzymic digestion of dAT copolymer with pancreatic DNase I to obtain fractions with degrees of polymerization from about 6 to beyond 40.
TL;DR: The phosphorylated intermediate in the (Na + K)-activated adenosine triphosphatase (Na-K ATPase) has been characterized as an L-glutamyl-γ-phosphate residue in the enzyme.
Abstract: The phosphorylated intermediate in the (Na + K)-activated adenosine triphosphatase (Na-K ATPase) has been characterized as an L-glutamyl-gamma-phosphate residue in the enzyme. This has been accomplished by digestion of the phosphorylated and nonphosphorylated forms of the enzyme with pepsin, reaction of the pepsin digests with [2,3-(3)H]N-(n-propyl)hydroxylmine, further digestion of the derivatized peptides with pronase in the presence of carrier L-glutamyl-gamma-N-(n-propyl)hydroxamate and carrier L-aspartyl-N-(n-propyl)hydroxamate, and chromatographic purification. An increment in radioactivity migrated with authentic L-glutamyl-gamma-N-(n-propyl)hydroxamate in a total of seven electrophoretic and chromatographic systems and on gel filtration. No increment in radioactivity was associated with authentic L-aspartyl-beta-N-(n-propyl)hydroxamate in five out of the seven chromatographic and electrophoretic systems. At the last stage of purification the radioactivity from the phosphorylated enzyme which migrated as L-glutamyl-gamma-N-(n-propyl)hydroxamate was 2(1/2) times that from the nonphosphorylated enzyme. On the basis of these results it is concluded that the phosphorylated intermediate in the Na-K ATPase is an L-glutamyl-gamma-phosphate residue. The beef brain Na-K ATPase has been solubilized with the nonionic detergent, Lubrol, and has been purified 10 times over that in the original microsomes. The soluble enzyme remains stable in the presence of ATP and either Na(+) or K(+). If the partially purified enzyme is electrophoresed in 3% polyacrylamide, followed by incubation with ATP, Na(+), K(+), and Mg(++), a single, somewhat diffuse, ATPase band, which is ouabain-sensitive is seen. Protein impurities are also seen on the gel. Gel electrophoresis, after treatment of the partially purified enzyme with phenol-acetic acid-urea, shows about 12 discrete protein bands. Studies on the site-directed alkylation of the (Na + K)-activated adenosine triphosphatase with haloacetate derivatives of cardiotonic steroids are reviewed. Efforts are now underway to specifically alkylate the cardiotonic steroid site of the Na-K ATPase with hellebrigenin 3-[2-(3)H]iodoacetate and to purify the subunit of the enzyme containing the cardiotonic steroid site by following radioactivity. Finally, a working model for the role of the Na-K ATPase in the coupled transport of Na and K is presented.
TL;DR: Comparison of the composition of the proteins showed that γ -A 2 differs from γ-B only in content of single residues of four amino acids and two substitutions, Ser/Arg and His/Pro are postulated, implying a close relationship in the synthesis of these milk proteins.
TL;DR: The resolution was greater with this procedure than with the continuous acrylamide gel technique, and the electrophoretic patterns were improved further by staining the gels with Coomassie Blue dye.
TL;DR: Response to inhibitors, heat-denaturation and competitive substrates suggests that a single active site is responsible for all four activities of the beta-glucosidase activity of pig kidney.
Abstract: 1. The β-glucosidase activity of pig kidney is located in the unsedimentable fraction of the cell and is not associated with the lysosomes. 2. The enzyme is active towards β-d-glucosides, β-d-galactosides, β-d-xylosides and α-l-arabinosides. 3. These activities could not be separated by gel electrophoresis, gel filtration or DEAE-cellulose chromatography. 4. Response to inhibitors, heat-denaturation and competitive substrates suggests that a single active site is responsible for all four activities. 5. Two forms of the enzyme were found to occur either separately or together in kidneys of pigs from several different breeds. 6. Electro-focusing experiments show these to have a small difference in isoelectric point (4·9 and 5·1), and gel filtration gives an approximate molecular weight of 50000 for both forms. 7. The characteristics of these two enzymes are compared.
TL;DR: It was found that the mobility of DNA on electrophoresis in agarose gel was inversely related to its sedimentation rate in sucrose density gradient, which has been commonly used to assess the relative molecular length of DNA.
TL;DR: Virus-specific antigens in cells infected with herpes simplex virus were separated on a preparative scale by two-stage polyacrylamide-gel electrophoresis by immunodiffusion and analytical Electrofiltration confirmed that separation had been achieved.
Abstract: Summary
Virus-specific antigens in cells infected with herpes simplex virus were separated on a preparative scale by two-stage polyacrylamide-gel electrophoresis. In the first stage, termed ‘electrofiltration’, the extract was electrophoresed through 3.5% gel to remove larger aggregates. Antigens in the resulting filtrate were then separated in the second stage by electrophoresis through 7% gel. The protein was recovered by sectioning the gel followed by overnight elution. Recoveries of about 70% were obtained from a 12 to 15 mg. load. Immunodiffusion and analytical electrophoresis both confirmed that separation had been achieved.
TL;DR: The PN-phosphorylase from vegetative cells was more anionic than that from the spores during gel electrophoresis in low concentrations of phosphate buffer, and the Stokes' radii and sedimentation constants of the vegetative cell enzyme were constant over a wide range of phosphate concentrations.