Absence of superoxide dismutase activity in a soluble cellular isoform of prion protein produced by baculovirus expression system

Sakudo, Akikazu; Hamaishi, M.; Hosokawa-Kanai, Tomoko; Tuchiya, Kotaro; Nishimura, Takuya; Saeki, Keiichi; Matsumoto, Yoshitsugu; Ueda, Seinosuke; Onodera, Takashi

doi:10.1016/s0006-291x(03)01239-7

Cited by 19 publications

(20 citation statements)

References 17 publications

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“…Assay for superoxide dismutase (SOD) activity. A SOD assay kit (WST Dojindo, Kumamoto, Japan) was used to measure the SOD activity as described previously (24,25,28,29). Each protein extract (20 µg) was assayed and compared with 1 U of bovine erythrocyte Cu/Zn-SOD activity.…”

Section: Mice Zrchi Prnpmentioning

confidence: 99%

Serum Withdrawal‐Induced Apoptosis in ZrchI Prion Protein (PrP) Gene‐Deficient Neuronal Cell Line Is Suppressed by PrP, Independent of Doppel

Nishimura

Sakudo

Hashiyama

et al. 2007

Microbiology and Immunology

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Previous studies have shown that cellular prion protein (PrPC) plays anti‐apoptotic and anti‐oxidative role against cell death induced by serum‐deprivation (SDP) in an immortalized prion protein gene‐deficient neuronal cell line derived from Rikn prion protein (PrP) gene‐deficient (Prnp–/–) mice, which ectopically produce excess Doppel (Dpl) (PrP‐like glycoprotein). To investigate whether PrPC inhibits apoptotic neuronal cell death without Dpl, an immortalized cell line was established from the brain of ZrchI Prnp–/– mice, which do not show ectopic expression of Dpl. The results using a ZrchI neuronal Prnp–/– cell line (NpL2) showed that PrPC potently inhibited SDP‐induced apoptotic cell death. Furthermore, PrPC expression enhanced the superoxide dismutase (SOD) activity in NpL2 cells. These results indicate that Dpl production did not affect anti‐apoptotic and anti‐oxidative functions of PrP, suggesting that PrPC may be directly correlated with protection against oxidative stress.

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Section: Mice Zrchi Prnpmentioning

confidence: 99%

Serum Withdrawal‐Induced Apoptosis in ZrchI Prion Protein (PrP) Gene‐Deficient Neuronal Cell Line Is Suppressed by PrP, Independent of Doppel

Nishimura

Sakudo

Hashiyama

et al. 2007

Microbiology and Immunology

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“…Immunoblotting of PrP C of baculovirus origin showed, in addition to the main band (mol. weight 27-29 kDa), additional high molecular-weight bands, various glycoforms [8], and low molecular-weight products, emerging as a result of partial proteolytic degradation [10]. A lower molecular weight of the baculovirus product in comparison with mol.…”

Section: Resultsmentioning

confidence: 99%

Isolation and characterization of full-length recombinant cattle PrPC protein

et al. 2006

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“…Further, it has been reported that Nagasaki-type PrP knockout mice showed an increase in heme oxygenase 1 and nitric oxide synthase levels, suggesting enhanced oxidative stress concomitant with Dpl expression (43). The proposal that PrP is a superoxide dismutase and therefore able to remove free radical species (44) would explain its beneficial effect in counterbalancing Dpl toxicity; however, this role of PrP is disputed (45)(46)(47). Conversely, it has been reported that PrP facilitates apoptosis (48,49).…”

Section: Discussionmentioning

confidence: 99%

Transgene-driven expression of the Doppel protein in Purkinje cells causes Purkinje cell degeneration and motor impairment

Anderson

Rossi

Linehan

et al. 2004

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

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The Doppel (Dpl) and Prion (PrP) proteins show 25% sequence identity and share several structural features with only minor differences. Dpl shows a PrP-like fold of its C-terminal globular domain and lacks the flexible N-terminal tail. The physiological functions of both proteins are unknown. However, ubiquitous Dpl overexpression in the brain of PrP knockout mice correlated with ataxia and Purkinje cell degeneration in the cerebellum. Interestingly, a similar phenotype was reported in transgenic mice expressing an N-terminally truncated PrP (⌬PrP) in Purkinje cells by the L7 promoter (TgL7-⌬PrP). Coexpression of full-length PrP rescued both the neurological syndromes caused by either Dpl or ⌬PrP. To evaluate whether the two proteins caused cerebellar neurodegeneration by the same mechanism, we generated transgenic mice selectively expressing Dpl in Purkinje cells by the same L7 promoter. Such mice showed ataxia and Purkinje cell loss that depended on the level of Dpl expression. Interestingly, the effects of high levels of Dpl were not counterbalanced by the presence of two Prnp alleles. By contrast, PrP coexpression was sufficient to abrogate motor impairment and to delay the neurodegenerative process caused by moderate level of Dpl. A similar situation was reported for the corresponding TgL7-⌬PrP mice supporting the concept that Dpl and ⌬PrP cause cell death, possibly by interfering with a common signaling cascade essential for cell survival.

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Absence of superoxide dismutase activity in a soluble cellular isoform of prion protein produced by baculovirus expression system

Cited by 19 publications

References 17 publications

Serum Withdrawal‐Induced Apoptosis in ZrchI Prion Protein (PrP) Gene‐Deficient Neuronal Cell Line Is Suppressed by PrP, Independent of Doppel

Serum Withdrawal‐Induced Apoptosis in ZrchI Prion Protein (PrP) Gene‐Deficient Neuronal Cell Line Is Suppressed by PrP, Independent of Doppel

Isolation and characterization of full-length recombinant cattle PrPC protein

Transgene-driven expression of the Doppel protein in Purkinje cells causes Purkinje cell degeneration and motor impairment

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