PMID: 8959022Oct 31, 1996Paper

Characterization of single-stranded cAMP response element binding protein (ssCRE-BP) from mouse cerebellum

Annals of the New York Academy of Sciences
T OsugiN Miki

Abstract

It has been speculated that opiate tolerance and dependence may occur at the level of gene expression. Our previous studies have shown that the binding activity of a nuclear factor (ssCRE-BP) to single-stranded CRE of somatostatin gene is altered by long-term treatment with morphine in the mouse cerebellum. ssCRE-BP was purified from the mouse cerebellum by a combination of chromatography on DNA affinity agarose and Mono Q HR. The native protein exhibited a molecular size of 110-150 kDa by gel filtration, and two polypeptides of about 35-40 kDa were observed on SDS-PAGE. The cloning and sequencing of a cDNA encoding ssCRE-BP showed that the protein possesses a glycine-rich domain and a glutamine-rich domain in the amino terminus and the carboxyl terminus, respectively. To investigate the function of ssCRE-BP in the brain, recombinant glutathione-S-transferase (GST) fusion proteins containing ssCRE-BP were expressed in bacterial systems. Rabbit anti-ssCRE-BP antibodies were raised against a GST-ssCRE-BP fusion protein. Using the antibodies in western blot analysis, a polypeptide of approximately 66 kDa was detected in the brain. These findings indicate that ssCRE-BP is involved in opiate tolerance and dependence.

References

May 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·T V NguyenS E Hyman
Sep 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·A M GraybielH A Robertson
Dec 15, 1988·Biochemical and Biophysical Research Communications·S L ChangR E Harlan
Mar 21, 1970·Nature·E Eidelberg, A S Schwartz
Jul 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·D DiMaioT Maniatis
Mar 1, 1981·Pharmacology, Biochemistry, and Behavior·L S Brady, S G Holtzman

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Citations

Jul 8, 2015·Electromagnetic Biology and Medicine·Christina L RossBenjamin S Harrison
Aug 4, 2004·Molecular and Biochemical Parasitology·Francisco Meirelles Bastos de OliveiraMarcelo Rosado Fantappié

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