Purification and Identification of a Human‐Serum DNA‐Binding Protein Associated with Malignant Diseases

Parsons, Robert G.; Hoch, James A.

doi:10.1111/j.1432-1033.1976.tb11082.x

Cited by 17 publications

(8 citation statements)

References 28 publications

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“…In addition to antibodies, other proteins have been found to react with DNA, such as C l q (31), C3DP (a fragment of C3 complement component) (42), and fibronectin (43). The last substance has been reported to play an important role in the removal of collagenous particles, fibrin and cell debris via sessile macrophages of the reticuloendothelial system (44,45).…”

Section: Discussionmentioning

confidence: 99%

Dna in synovial fluid and the circulation of patients with arthritis

Leon

Revach²,

Ehrlich

et al. 1981

Arthritis & Rheumatism

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DNA levels were measured in synovial fluids and sera of 106 patients with rheumatoid arthritis (RA), osteoarthritis (OA), gout, pseudogout, and posttraumatic arthritis (TRA). In synovial fluids, the highest concentration was found in rheumatoid arthritis (mean f SE 18 f 3 pg/d for seropositive and 9 f 1 pg/d for seronegative variants), gout and pseudogout (17 f 3 pg/ ml). In contrast, the levels in patients with OA or acute TRA were very low: 0.8 f 0.1 pg/d and 1.1 f 0.2 pg/ ml, respectively. The difference between the means of the first disease group and OA or TRA is statistically significant. A similar pattern was observed for DNA levels in the circulation: in rheumatoid arthritis, the mean concentration was 135 f 28 ng/ml and 164 f 39 n g / d for seropositive and seronegative RA, respectively. Again the levels in OA and TRA were much lower, 52 k 18 ng/ml and 0 ng/ml, respectively. The latter are not significantly different from the mean levels of 95 normal,

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Section: Discussionmentioning

confidence: 99%

Dna in synovial fluid and the circulation of patients with arthritis

Leon

Revach²,

Ehrlich

et al. 1981

Arthritis & Rheumatism

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“…Introduction DNA-binding proteins are known to be present in human serum and some have been described [ 1,2] . Their biological significance is unknown and the relationship between serum and intra-nuclear DNAbinding proteins is obscure although the latter may be related to the control of gene expression.…”

mentioning

confidence: 99%

“…Their biological significance is unknown and the relationship between serum and intra-nuclear DNAbinding proteins is obscure although the latter may be related to the control of gene expression. The appearance of carcinofetal antigens such as CEA [3] , alphafetoprotein [4] and the DNA-binding protein C3DP [2] in malignant disease may indeed be the result of derepression of the original foetal genome. A unique 36 000 dalton DNA-binding protein in foetal cord and malignant disease serum has been reported [5] .…”

mentioning

confidence: 99%

A serum DNA‐binding protein absent in malignant disease

Lewis

André

1978

FEBS Letters

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“…molecules, e.g., the 26,000-dalton nuclear antigen described by Yeoman et al (22) or the DNA binding proteins isolated from human sera (23,24).…”

Section: Discussionmentioning

confidence: 99%

Purification of nuclear antigens in Novikoff hepatoma.

Fujitani¹,

Chiu²,

Hnilica³

1978

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

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Nuclear antigen in Novikoff hepatoma chromatin was partially purified and characterized. As indicated b complement fixation assay, this antigen was present in chromatin of embryonic livers and several transplantable tumors. It was not detected in normal tissue chromatins of the same animals. For its immunological specificity this protein antigen (molecular weight 45,000-0,000) had to be complexed with DNA. Preliminary experiments indicate that specific nuclear protein antigens are also present in human tissues and spontaneous malignancies. Nuclear transplants as well as other experiments indicate that malignant transformation may reflect a profound change in cellular differentiation. The new (cancerous) phenotype of the newly differentiated cells is maintained through the cellular progenies during the proliferation of the tumor. In view of the widely accepted notion that certain chromosomal nonhistone proteins may be responsible for the tissue-specific regulation of genetic expression, the identification of such regulatory proteins associated with malignant growth is a necessary step toward the basic understanding of carcinogenic mechanism at the level of gene transcription. Assuming that the regulatory proteins may be specific for the particular types of cells and tissues, we have used an immunological approach in the search for such macromolecules. We report here a partial purification of a growth-associated nuclear antigen in Novikoff hepatoma. MATERIALS AND METHODSIsolation of Nuclei and Chromatin. Rat liver nuclei were isolated by a modification of the hypertonic sucrose procedure of Blobel and Potter (1). Novikoff hepatoma nuclei were isolated by hypotonic shock, high pressure shearing, and centrifugation in hypertonic sucrose (2). The isolation of chromatin was described previously (3).Fractionation of Chromosomal Proteins. For immunization, isolated chromatins were dehistonized by dissociation with 5 M urea and 2.5 M NaCl in 50 mM sodium succinate buffer (pH 5.0) (4). After ultracentrifugation, the pellets containing chromosomal nonhistone proteins and DNA were homogenized in 0.1 mM MgGI2/2 mM Tris-HCl buffer (pH 7.5). The suspension was stirred overnight at 40 and centrifuged at 1000 X g for 5 min to sediment a small amount of particulate material. The supernatant was used for immunization.For fractionation by hydroxylapatite chromatography, the bulk of chromosomal nonhistone proteins was first removed by extraction with 5.0 M urea/0. 1 mM phenylmethylsulfonyl fluoride in 50 mM sodium phosphate buffer (pH 7.6). The mixture was gently homogenized (0.3-0.5 mg of DNA per ml) and stirred for 2 hr in an ice bath. Chromatin, partially deproteinized by this procedure (removal of nearly 50% of total chromatin protein content), was sedimented by centrifugation at 15,000 X g for 60 min. The pellet containing DNA, histones, and a small amount of nonhistone proteins was rehydrated by homogenization in 1.5 mM NaCl/0.15 mM sodium citrate/0.1 mM phenylmethylsulfonyl fluoride at a concentration of 2 mg of DNA ...

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Purification and Identification of a Human‐Serum DNA‐Binding Protein Associated with Malignant Diseases

Cited by 17 publications

References 28 publications

Dna in synovial fluid and the circulation of patients with arthritis

Dna in synovial fluid and the circulation of patients with arthritis

A serum DNA‐binding protein absent in malignant disease

Purification of nuclear antigens in Novikoff hepatoma.

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