The homology of a novel polypeptide with stress protein characteristics in embryonic mice brain and in the hypothalamus of caloric restricted rats as determined by ultramicro western blotting

Pipkin, James L.; Hinson, William G.; Lyn‐Cook, Lascelles E.; Borde, James La; Anson, Jeanne F.; Feuers, Ritchie J.; Duffy, Peter H.; Hart, Ronald W.; Casciano, Daniel A.

doi:10.1002/elps.1150150196

Cited by 10 publications

(1 citation statement)

References 39 publications

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“…Proteins were identified as previously described [29]. Individual sps (sp25a, 70i, 72c, 90) were extracted electrophoretically from gel plugs taken from the second-dimensional slabs and extracted in an ISCO protein isolation-concentrator "little blue tank" (ISCO, Lincoln, NE, USA) [21,30]. SDS was removed from the proteins during extraction [21,31].…”

Section: Electrophoresis and Extraction Of Stress Proteins From Gelsmentioning

confidence: 99%

Decreasedin vitro interaction between p53 and nuclear stress proteins in thep53-deficient mouse

Morris¹,

Pipkin²,

Hinson³

et al. 2001

Electrophoresis

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In a previous study, the strength of the interaction between the nuclear stress proteins (sps) 25a, 70i, 72c, and 90 and the tumor suppressor protein p53 was determined by an in vitro fluorescence binding assay. The relative binding of the individual sps with p53, derived from the bone marrow of transgenic mice heterozygous at the p53 locus (p53+/-), was reduced compared to the interaction of sps and p53 derived from wild-type (p53+/+) mice. In order to determine if the genotype of the p53 donor or the genotype of the sp donor determined the binding efficiency, p53 expression was induced by retinoic acid and sp synthesis by bleomycin. P53 derived from either wild-type or heterozygous animals was cross-reacted with nuclear sps obtained from either wild-type or heterozygous animals. Each of the sps, 25a, 70i, 72c, and 90, bound to wild-type p53 with a similar efficiency, irrespective of the genotype of the sp donor mouse (p53+/+ or p53+/-). In contrast, when the sp interaction with p53 obtained from the heterozygous mouse was measured, the relative value of the fluorescence complex was significantly reduced. The data suggest that the strength of the interaction between p53 and nuclear sps is related to the genotype of the p53 donor, and not to the genotype of the animals from which the sps are derived.

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Section: Electrophoresis and Extraction Of Stress Proteins From Gelsmentioning

confidence: 99%

Decreasedin vitro interaction between p53 and nuclear stress proteins in thep53-deficient mouse

Morris¹,

Pipkin²,

Hinson³

et al. 2001

Electrophoresis

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Induction of stress proteins by electromagnetic fields in cultured HL-60 cells

Pipkin

Hinson

Young

et al. 1999

Bioelectromagnetics

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HL‐60 cells in culture were exposed for 2 h to a sinusoidal 0.1 or 1 mT (1 or 10 Gauss) magnetic field at 60 Hz and pulse labeled after exposure with radioactive isotopes by incubation by using either [35S]methionine, [3H]leucine, or [33P]phosphate. The radioactive labels were incorporated into cellular proteins through synthesis or phosphorylation. Proteins were extracted from electrostatically sorted nuclei, and the heat shock/stress proteins (sp) were analyzed for synthesis and phosphorylation by two‐dimensional polyacrylamide gel electrophoresis. In the control cultures (no exposure to the magnetic field), sp 72c (cognate form) was faintly observed. A 0.1 mT exposure did not show sp metabolism to be different from that of the controls; however, after a 1 mT exposure of the HL‐60 cells, sp 70i (inducible form) was synthesized ([35S]methionine incorporation). Sp 90 was not synthesized at either field level, but was phosphorylated ([33P]phosphate incorporation) in the 1 mT exposure. Sp 27 (isoforms a and b) was induced after a 1 mT exposure as reflected by labeling with [3H]leucine. These sps were not detected after a 0.1 mT exposure. After a 1 mT exposure and labeling with [33P], sp 27 isoforms b and c were phosphorylated whereas isoform ‘a‘ was not observed. Sps 70i, 72c, and 90 were identified by commercial sp antibodies. Likewise, polypeptides a, b, and c were verified as sp 27 isoforms by Western blotting. Statistical evaluation of sp areas and densities, determined from fluorographs by Western‐blot analysis, revealed a significant increase in sps 90 and 27a after a 1 mT magnetic field exposure. The 1 mT magnetic field interacts at the cellular level to induce a variety of sp species. Bioelectromagnetics 20:347–357, 1999. Published 1999 Wiley‐Liss, Inc.

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The effect of aging and dietary restriction on the retinoylation of nuclear matrix proteins in rats

Pipkin

Hinson²,

Lyn‐Cook

et al. 1996

Aging Clin Exp Res

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The homology of a novel polypeptide with stress protein characteristics in embryonic mice brain and in the hypothalamus of caloric restricted rats as determined by ultramicro western blotting

Cited by 10 publications

References 39 publications

Decreasedin vitro interaction between p53 and nuclear stress proteins in thep53-deficient mouse

Decreasedin vitro interaction between p53 and nuclear stress proteins in thep53-deficient mouse

Induction of stress proteins by electromagnetic fields in cultured HL-60 cells

The effect of aging and dietary restriction on the retinoylation of nuclear matrix proteins in rats

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