PMID: 9003076Feb 1, 1997Paper

Stably transfected human cells overexpressing rat brain endopeptidase 3.4.24.16: biochemical characterization of the activity and expression of soluble and membrane-associated counterparts

Journal of Neurochemistry
B VincentF Checler

Abstract

We recently cloned endopeptidase-24.16 (neurolysin; EC 3.4.24.16), a neurotensin-degrading peptidase likely involved in the physiological termination of the neurotensinergic signal in the central nervous system and in the gastrointestinal tract. We stably transfected human kidney cells with the pcDNA3-lambda 7aB1 construction bearing the whole open reading frame encoding the rat brain peptidase. Transfectants displayed endopeptidase-24.16 immunoreactivity and exhibited QFS- and neurotensin-hydrolyzing activities, the biochemical and specificity properties of which fully matched those observed with the purified murine enzyme. Cryoprotection experiments and substrate degradation by intact plated cells indicated that transfectants exhibited a membrane-associated form of endopeptidase-24.16, the catalytic site of which clearly faced the extracellular domain. Transfected cells were unable to secrete the enzyme. Overall, our experiments indicate that we have obtained stably transfectant cells that overexpress an enzymatic activity displaying biochemical properties identical to those of purified endopeptidase-24.16. The membrane-associated counterpart and lack of secretion of the enzyme were clearly reminiscent of what was observed wi...Continue Reading

Citations

Sep 21, 2007·Current Eye Research·Rubens Bertazolli-FilhoEduardo Miguel Laicine
Mar 7, 2006·Journal of Neurochemistry·Nathaniel A JeskeJames L Roberts
Mar 1, 2003·American Journal of Physiology. Heart and Circulatory Physiology·M Ursula NormanRebecca A Lew
Oct 10, 2002·Endocrine Reviews·Corie N ShrimptonRebecca A Lew
Aug 31, 2018·Neurochemical Research·Frédéric Checler, Emer S Ferro
Sep 18, 2001·The Journal of Comparative Neurology·J D Fontenele-NetoE S Ferro
Aug 23, 2002·Protein Science : a Publication of the Protein Society·Kallol RayDavid W Rodgers
Oct 16, 2007·Bioorganic & Medicinal Chemistry Letters·Jennifer K MitchellRonald J Quinn

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