| Title: | Cleavage-secretion of angiotensin I-converting enzyme in yeast |
| Author(s): | Williams TA; Gouttaya M; Tougard C; Michaud A; Chauvet MT; Corvol P; |
| Address: | "INSERM unite 36, College de France, Paris. williams@infobiogen.fr" |
| DOI: | 10.1016/s0303-7207(97)04022-7 |
| ISSN/ISBN: | 0303-7207 (Print) 0303-7207 (Linking) |
| Abstract: | "Angiotensin I-converting enzyme (ACE) is a type I transmembrane protein composed of two domains (N and C domains) which undergoes a post-translational proteolytic cleavage in mammalian cells to release the soluble ectodomain. The protease involved in ACE cleavage-secretion (ACE-secretase) is not well characterised and eludes isolation: the presence of a yeast homologue, thus more amenable to genetic manipulation, would facilitate its identification. We have expressed a secreted form of the ACE C domain, lacking the C-terminal membrane anchor (C domain(deltaCOOH)), and the membrane-anchored C domain (C domain) in the yeast Pichia pastoris by fusion to prepro-alpha-factor. Immunofluorescent labelling localises the ACE C domain to the periphery of yeast cells but not C domain(deltaCOOH), however, expression of both C domain and C domain(deltaCOOH) produced soluble enzymes in the culture medium. Immunocharacterisation of the two soluble forms of the C domain indicates a proteolytic cleavage of the membrane-bound C domain to produce the soluble counterpart. Thus ACE undergoes a proteolytic cleavage in yeast" |
| Keywords: | "Amino Acid Sequence Cloning, Molecular Endopeptidases/*metabolism Humans Kinetics Mating Factor Molecular Sequence Data Mutagenesis, Site-Directed Peptide Biosynthesis *Peptides Peptidyl-Dipeptidase A/biosynthesis/chemistry/*metabolism Pichia Protease Inh;" |
| Notes: | "MedlineWilliams, T A Gouttaya, M Tougard, C Michaud, A Chauvet, M T Corvol, P eng Research Support, Non-U.S. Gov't Ireland 1997/04/04 Mol Cell Endocrinol. 1997 Apr 4; 128(1-2):39-45. doi: 10.1016/s0303-7207(97)04022-7" |
