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Volume 271, Number 28, Issue of July 12, 1996 pp. 16856-16861
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Effect of Scrapie Infection on the Activity of Neuronal Nitric-oxide Synthase in Brain and Neuroblastoma Cells

(Received for publication, October 24, 1995, and in revised form, February 20, 1996)

From the Department of Neurology, Hadassah Hebrew University Hospital, Jerusalem, Israel 91120

Nitric-oxide synthase (NOS) is responsible for the synthesis of nitric oxide which serves as a neural messenger in the central nervous system. NOS activity was markedly inhibited in brains of mice and hamsters and neuroblastoma cells infected with scrapie (ScN2a). The decrease in activity was in accordance with decreased NADPH-diaphorase-positive cells and decreased staining of NOS-positive cells demonstrated by specific anti-NOS antibodies. However, the specific nNOS mRNA in ScN2a was elevated when compared with normal neuroblastoma cells (N2a). Immunoblotting of fractions from these cell lines with an anti-nNOS monoclonal antibody revealed a band of nNOS from N2a and two bands with a lower molecular weight in ScN2a cells. Furthermore, NOS in ScN2a cells was insoluble in nondenaturing detergents. This insolubility is one of the landmark properties of PrP^Sc. It is, therefore, possible that nNOS in scrapie-infected cells and brains is aberrantly folded, resulting in an insoluble and inactive enzyme.

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