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Published online on January 27, 2003, 10.1073/pnas.0335507100
PNAS | February 4, 2003 | vol. 100 | no. 3 | 928-933


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Biochemistry
 The secretory proprotein convertase neural apoptosis-regulated convertase 1 (NARC-1): Liver regeneration and neuronal differentiation

(cleavage specificity|hypercholesterolemia|neurogenesis|hepatogenesis)

Nabil G. Seidah*,dagger , Suzanne Benjannet*, Louise Wickham*, Jadwiga Marcinkiewicz*, Stéphanie Bélanger JasminDagger , Stefano StifaniDagger , Ajoy Basak§, Annik Prat*, and Michel Chrétien§

* Laboratory of Biochemical Neuroendocrinology, Clinical Research Institute of Montreal, 110 Pine Avenue West, Montreal, QC, H2W 1R7 Canada; Dagger  Montreal Neurological Institute, McGill University, Montreal, QC, H3A 2B4 Canada; and § Regional Protein Chemistry Center and Diseases of Aging Unit, Ottawa Health Research Institute, Ottawa Hospital, Civic Campus, 725 Parkdale Avenue, Ottawa, ON, K1Y 4E9 Canada

Edited by Donald F. Steiner, University of Chicago, Chicago, IL, and approved December 5, 2002 (received for review September 10, 2002)

Seven secretory mammalian kexin-like subtilases have been identified that cleave a variety of precursor proteins at monobasic and dibasic residues. The recently characterized pyrolysin-like subtilase SKI-1 cleaves proproteins at nonbasic residues. In this work we describe the properties of a proteinase K-like subtilase, neural apoptosis-regulated convertase 1 (NARC-1), representing the ninth member of the secretory subtilase family. Biosynthetic and microsequencing analyses of WT and mutant enzyme revealed that human and mouse pro-NARC-1 are autocatalytically and intramolecularly processed into NARC-1 at the (Y,I)VV(V,L)(L,M)down-arrow motif, a site that is representative of its enzymic specificity. In vitro peptide processing studies and/or Ala substitutions of the P1-P5 sites suggested that hydrophobic/aliphatic residues are more critical at P1, P3, and P5 than at P2 or P4. NARC-1 expression is highest in neuroepithelioma SK-N-MCIXC, hepatic BRL-3A, and in colon carcinoma LoVo-C5 cell lines. In situ hybridization and Northern blot analyses of NARC-1 expression during development in the adult and after partial hepatectomy revealed that it is expressed in cells that have the capacity to proliferate and differentiate. These include hepatocytes, kidney mesenchymal cells, intestinal ileum, and colon epithelia as well as embryonic brain telencephalon neurons. Accordingly, transfection of NARC-1 in primary cultures of embryonic day 13.5 telencephalon cells led to enhanced recruitment of undifferentiated neural progenitor cells into the neuronal lineage, suggesting that NARC-1 is implicated in the differentiation of cortical neurons.

dagger To whom correspondence should be addressed. E-mail: seidahn{at}ircm.qc.ca.

www.pnas.org/cgi/doi/10.1073/pnas.0335507100
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