Purification and Characterization of a Bovine Cerebral Cortex Cell Surface Sialoglycopeptide that Inhibits Cell Proliferation and Metabolism

Sharifi, Behrooz G.; Johnson, Terry C.; Khurana, Vijay K.; Bascom, Charles C.; Fleenor, T. J.; Chou, Hsin-Hwei

doi:10.1111/j.1471-4159.1986.tb12990.x

Cited by 44 publications

(29 citation statements)

References 31 publications

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“…The antibody was radioiodinated by the chloramine-T method of Greenwood et al (1963) with the modifications described by Sharifi et al (1986).…”

Section: Methodsmentioning

confidence: 99%

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Reconstitution with T Lymphocytes Protects Nude Mice from a Central Nervous System Disorder Induced by a Temperature-sensitive Vesicular Stomatitis Virus

Doll

Johnson²

1988

Journal of General Virology

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SUMMARYA temperature-sensitive mutant of vesicular stomatitis virus (VSV), tsG31-KS5 VSV, intracerebrally inoculated into BALB/c (+/+) or Swiss outbred mice yielded a clinically asymptomatic persistent infection of the central nervous system (CNS). BALB/c nude (nu/nu) mice infected with tsG31-KS5 VSV, however, all perished within 26 days of infection. All the nude mice were afflicted with a slowly progressing CNS disorder, with symptoms including lethargy, curvature of the spine, hind-limb paralysis and other neurological disorders, before they succumbed to the infection. Wild-type (wt) VSV infection of either normal or nude mice, on the other hand, invoked a rapidly lethal disease with all animals dying within 4 days of infection. When nude mice were reconstituted with 5 x 106 syngeneic T lymphocyte-enriched splenocytes, over 70% of them not only survived the tsG31-KS5 VSV infection but appeared to be free of any neurological disorders. Only 20 % of these reconstituted mice infected for 20 days with tsG31-KS5 VSV endured a wt VSV challenge. In contrast, BALB/c (+/+) mice infected for 20 days with tsG31-KS5

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“…The antibody was radioiodinated by the chloramine-T method of Greenwood et al (1963) with the modifications described by Sharifi et al (1986).…”

Section: Methodsmentioning

confidence: 99%

“…Solid phase radioimmunoassays (SPRIA) were performed as described previously (Sharifi et al, 1986), with the following modifications. The wells of 96-well microtitre plates were coated with tsG31-KS5 VSV [20 p.g/ml in phosphate-buffered saline (PBS)] at 4 °C overnight.…”

Section: Methodsmentioning

confidence: 99%

Reconstitution with T Lymphocytes Protects Nude Mice from a Central Nervous System Disorder Induced by a Temperature-sensitive Vesicular Stomatitis Virus

Doll

Johnson²

1988

Journal of General Virology

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“…The gel permeation HPLC purified sample was resuspended in 0.1 M phosphate buffer, 10% glycerol, and 55% ammonium sulfate. In some cases the sample was further purified by hydrophobic interaction HPLC with a phenyl hydrophobic column with a linear gradient of a decreasing concentration of ammonium sulfate [9]. The fraction that contained inhibitory activity was dialyzed and concentrated and stored at -70°C until further study.…”

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confidence: 99%

Relationship between protease activity and a sialoglycopeptide inhibitor isolated from bovine brain

Sharifi

Bascom

Fattaey

et al. 1986

J of Cellular Biochemistry

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We have recently described the isolation and purification to homogeneity of a new sialoglycopeptide from bovine brain cell surfaces that reversibly inhibits protein synthesis and DNA synthesis of normal but not transformed cells. Active inhibitory preparations, however, were shown to contain a protease activity that was not lost upon purification. Several experiments were performed to establish the relationship between the proteolytic activity of the sialoglycopeptide and the biological inhibitory activity. Both the protease activity and inhibitory activity were stable at pH 6-8 but were reduced or completely destroyed below pH 4 and above pH 9. Acid inactivation was reversible and upon dialysis, both the biological inhibitory and protease activities were regained. Deglycosylation and CNBr cleavage indicated that the polypeptide backbone, rather than carbohydrate moiety, played an important role in the protease and biological inhibitory activities. Furthermore, chemical modification of amino and tyrosine groups indicated that both residues are essential for both activities. Thus, the biological inhibitory activity and protease activity are very closely related and most likely reside with the same polypeptide sequence.

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“…A sialoglycopeptide (SGP) from bovine brain (molecular weight of 18 kDa) was reported to inhibit cell growth ofafibroblast cell line, NIH 3T3 cells (4,13). Inhibition of DNAsynthesis was shown in various cells; fibroblast cell, tumor cell, transformed cell and also primary cultured cell (4).…”

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confidence: 99%

“…The effect of SGP on the PC12 cell growth was analyzed using the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay (1, ll Preparation of the partially purified SGP. The SGP was prepared according to the method of Kinders et al (9,13). Bovine brains, obtained from a local slaughter house, were immediately frozen and stored at -20°C.…”

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Clarification of PC12 Cell Growth Inhibition by a Sialoglycopeptide from Bovine Brain under Low Serum Condition.

Kobayashi

Shinozawa

1994

Cell Struct. Funct.

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ABSTRACT. To study the effect of a bovine brain sialoglycopeptide (SGP) on the nerve cell growth, PC12 cells were used as a model system. Though the inhibition of cell growth, measured by the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay, by SGP was slight under the serum concentration of 10%, it was clearly detected under that of 1 to 2%. For the preparation of SGP, filtration through a nitrocellulose membranewas useful.A sialoglycopeptide (SGP) from bovine brain (molecular weight of 18 kDa) was reported to inhibit cell growth ofafibroblast cell line, NIH 3T3 cells (4, 13). Inhibition of DNAsynthesis was shown in various cells; fibroblast cell, tumor cell, transformed cell and also primary cultured cell (4). The presence of the SGP receptors in 3T3 cells was shown by use of the 125I-labeled SGP (2, 14). The native form of SGP with molecular weight of 66,000 was isolated from the cell membrane in bovine brain (5). The above data suggest that the SGPmight participate in the contact inhibition in these cells.In this paper, the PC12 cell, rat pheochromocytoma, was used as a model of premature neuron, having the potential of a neuron-like outgrowth whenexposed to NGF(6). The effect of SGP on the PC12 cell growth was analyzed using the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay (1, ll). The MTT assay is based on the dehydrogenase activity in mitochondria of living cells. Therefore, this assay is used to determine the living cell number. The inhibitory effect of SGPunder 10%serum condition was not clearly detected. Whenthe serum concentration in the media was decreased to the range of 1 to 2%, the effect was clearly detected. This dependency on the serum concentration probably originated from the presence of cell growth factors (12) and also inhibitory factors (3, 7) in the serum. MATERIALS AND METHODSChemicals. Materials were obtained from the following sources: Dulbecco's modified Eagle's medium(DMEM) and F12 nutrient mixtures (Ham (F12)) (Gibco); fetal bovine serum (FBS) and horse serum (HS) (Flow Laboratory); bovine serum albumin, pronase E (from Streptomyces griseus), MTT (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide) and polyethylenimine (Sigma); SDS-PAGElow molecular weight marker (Pharmacia); nitrocellulose membrane filter (pore size 0.2 //m) for the sterilization of the culture medium or SGP solution, plates and dishes for cell culture (Corning). All other chemicals were reagent-grade materials from commercial sources.

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Purification and Characterization of a Bovine Cerebral Cortex Cell Surface Sialoglycopeptide that Inhibits Cell Proliferation and Metabolism

Cited by 44 publications

References 31 publications

Reconstitution with T Lymphocytes Protects Nude Mice from a Central Nervous System Disorder Induced by a Temperature-sensitive Vesicular Stomatitis Virus

Reconstitution with T Lymphocytes Protects Nude Mice from a Central Nervous System Disorder Induced by a Temperature-sensitive Vesicular Stomatitis Virus

Relationship between protease activity and a sialoglycopeptide inhibitor isolated from bovine brain

Clarification of PC12 Cell Growth Inhibition by a Sialoglycopeptide from Bovine Brain under Low Serum Condition.

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