Identification and isolation of a 155-kDa protein with neuropathy target esterase activity. NLM AIDSLINE Important note: Information in this article was accurate in 1997. The state of the art may have changed since the publication date.

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Identification and isolation of a 155-kDa protein with neuropathy target esterase activity.

Fundam Appl Toxicol. 1996 Mar;30(1):23-30. Unique Identifier : AIDSLINE MED/96436320
Mackay CE; Hammock BD; Wilson BW; Department of Avian Science and Environmental Toxicology, University; of California, Davis 95616, USA.

Abstract: A method is presented for the isolation of a 155-kDa protein that possesses phenyl valerate hydrolysis activity in the presence of paraoxon but is inhibited by mipafox; the functional definition of neuropathy target esterase (neurotoxic esterase; NTE). Microsomes, isolated from 18-day-old chicken embryos were treated with phospholipase A2 to solubilize the NTE activity. The extract was then combined with polyoxyethylene W1 detergent and resolved by gel filtration chromatography to yield an active fraction with an approximate mass of 200 kDa. This fraction was further purified by preparative isoelectric focusing and native electrophoresis to yield two separate bands possessing NTE activity. The slower migrating band was highly enriched in a 155-kDa protein that was identified as a source of the NTE activity by affinity chromatography using 3-(9'-mercaptononylthio)-1,1,1-trifluoro-propan-2-one bound to Sepharose CL6B. This represents the first report of the isolation of NTE in its active form and aids in the confirmation of the 155-kDa protein as the most likely candidate for NTE.
Keywords: *Carboxylic Ester Hydrolases/ANALYSIS *Carboxylic Ester Hydrolases/ISOLATION & PURIF *Proteins/ISOLATION & PURIF *Proteins/PHYSIOLOGYKWDcarboxylicesterhydrolases/analysisKWDcarboxylicesterhydrolases/isolation&purifKWDproteins/isolation&purifKWDproteins/physiology
970430
M9741536

Copyright © 1997 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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