Chromatography of Mixed Oligonucleotides on DEAE-Sephadex

Rushizky, George W.; Bartos, Edwin M.; Sober, Herbert A.

doi:10.1021/bi00893a005

Cited by 94 publications

(33 citation statements)

References 16 publications

(11 reference statements)

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“…The snake venom phosphodiesterase hydrolysate was mixed with urea at a final concentration of 7 M and applied to a DEAE-Sephadex A-25 column (0.5 X 70 cm) equilibrated with 50 mM Tris-HCI, pH 7.5/7 M urea (20). The peak fractions were designated as peaks A, B, and C (Fig.…”

mentioning

confidence: 99%

Structure of poly(adenosine diphosphate ribose): identification of 2'-[1''-ribosyl-2''-(or 3''-)(1'''-ribosyl)]adenosine-5',5'',5'''-tris(phosphate) as a branch linkage.

Misawa¹,

Saikawa²,

Yamaizumi³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

138

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Poly([14C]adenosine diphosphate ribose) was synthesized from [14C]NAD+ with calf thymus nuclei. The fraction containing poly(adenosine diphosphate ribose) eluted with 0.22--0.40 M phosphate buffer (pH 6.8) from a hydroxylapatite column, was completely hydrolyzed with venom phosphodiesterase, and was separated by DEAE-Sephadex A-25 column chromatography in 7 M urea. A new compound, which constituted 2% of the products from poly(adenosine diphosphate ribose), was found in addition to the expected products--i.e., 5'-AMP, 2'-(1''-ribosyl)adenosine-5',5''-bis(phosphate), and its derivatives. This compound was identified as 2'-[1''-ribosyl 2''-(or 3''-)(1'''-ribosyl)]adenosine-5',5'',5'''-tris(phosphate). The existence of this compound is evidence of a branching structure of poly(adenosine diphosphate ribose), which was previously thought to be a linear molecule. The content of this compound suggests that the frequency of branching is about 1 per 20--30 adenosine diphosphate ribose residues of high molecular weight poly(adenosine diphosphate ribose).

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mentioning

confidence: 99%

Structure of poly(adenosine diphosphate ribose): identification of 2'-[1''-ribosyl-2''-(or 3''-)(1'''-ribosyl)]adenosine-5',5'',5'''-tris(phosphate) as a branch linkage.

Misawa¹,

Saikawa²,

Yamaizumi³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

138

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“…The reaction mixture was incubated at 37 9 C for 3 or 6 h, and the reaction was stopped by the addition of urea to give a final concentration of 7 M. The fractionation of mononucleotides from the digest was done by the method of Rushizky et al 16 ) Mononucleotides were analyzed by HPLC on a Shim-pack WAX-I column in a Shimadzu LC-9A apparatus (Kyoto, Japan) at room temperature. The solvent was 20mM phosphate buffer, pH 7.0.…”

Section: Identification Of 5'-mononucleotides In Hydrolysate Of Yeastmentioning

confidence: 99%

Purification and Some Properties of Plant Endonuclease from Scallion Bulbs

Uchida

Takadera

et al. 1993

Bioscience, Biotechnology, and Biochemistry

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“…Among oligonucleotides, those up to trinucleotide can be dialyzed by use of cellophane tubing and therefore the molecular size of DF can be estimated to be less than 3,000. According to the elution profile of DEAE-Sephadex A-25 chromatography, the chain length of DF is situated between mononucleotide and trinucleotide, because at pH 7.5, where each of the nucleotide residues has the same negative charge due to the phosphate groups, separation of nucleotides occurs as a function of chain length in this buffer system (23).…”

Section: Ion Exchange Chromatography Of Dfmentioning

confidence: 99%

Immunogenic Dialyzable Factor Derived from a Ribosomal Fraction of Salmonella typhimurium

Kita

Matsuura²,

Masuda

et al. 1983

Microbiology and Immunology

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The preparation, properties, and immunogenicity of the dialyzable factor from a ribosomal fraction of Salmonella typhimurium are described. The ribosomal fraction was purified to eliminate O-antigenic components, by affinity chromatography (Sepharose-anti-O antibody conjugates used as immunoadsorbent). The dialyzable factor was obtained in the concentrated dialysate of the purified ribosomal fraction which was alternately frozen in dry-ice acetone and thawed in an 80 C water bath, for a total of five or six cycles. When this preparation was tested for its ability to protect mice against challenge with 1,000 LD 50 of the homologous bacteria, it afforded 100% protection at a dose equivalent to 5.0 pg of RNA. The protection conferred by this factor was mainly cell mediated but immune serum enhanced this immunity despite the fact that no antibodies were detected in it. The protective activity of this factor was sensitive to RNase digestion but resistant to proteolytic enzymes. Ion exchange chromatography of this factor with DEAE-Sephadex A-25 (in 7 M Urea-0.02 M Tris-HCI buffer, pH 7.5) resulted in a single A260 peak which was found to be immunogenic. Chemical analysis of this peak after it was concentrated and desalted revealed that this immunogenic fraction was composed mainly of mixed nucleotides. The data indicate that protective immunity conferred by a ribosomal vaccine is associated with RNA but may not require the intact RNA molecule.The immunizing efficacy of a bacterial ribosomal preparation was first reported in the case of Mycobacterium tuberculosis (34). Since then, ribosomal vaccines from a number of organisms have been shown to confer protection against challenge with the homologous organisms (6). However, in most instances these preparations ",Present address:

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Chromatography of Mixed Oligonucleotides on DEAE-Sephadex

Cited by 94 publications

References 16 publications

Structure of poly(adenosine diphosphate ribose): identification of 2'-[1''-ribosyl-2''-(or 3''-)(1'''-ribosyl)]adenosine-5',5'',5'''-tris(phosphate) as a branch linkage.

Structure of poly(adenosine diphosphate ribose): identification of 2'-[1''-ribosyl-2''-(or 3''-)(1'''-ribosyl)]adenosine-5',5'',5'''-tris(phosphate) as a branch linkage.

Purification and Some Properties of Plant Endonuclease from Scallion Bulbs

Immunogenic Dialyzable Factor Derived from a Ribosomal Fraction of Salmonella typhimurium

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