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• W1966307736 endingPage "185" @default.
• W1966307736 startingPage "185" @default.
• W1966307736 abstract "Angiotensin-converting enzyme (ACE) is one of a growing number of integral membrane proteins that is shed from the cell surface through proteolytic cleavage by a secretase. To investigate the requirements for ectodomain shedding, we replaced the glycosylphosphatidylinositol addition sequence in membrane dipeptidase (MDP) - a membrane protein that is not shed - with the juxtamembrane stalk, transmembrane (TM) and cytosolic domains of ACE. The resulting construct, MDP-STM(ACE), was targeted to the cell surface in a glycosylated and enzymically active form, and was shed into the medium. The site of cleavage in MDP-STM(ACE) was identified by MS as the Arg(374)-Ser(375) bond, corresponding to the Arg(1203)-Ser(1204) secretase cleavage site in somatic ACE. The release of MDP-STM(ACE) and ACE from the cells was inhibited in an identical manner by batimastat and two other hydroxamic acid-based zinc metallosecretase inhibitors. In contrast, a construct lacking the juxtamembrane stalk, MDP-TM(ACE), although expressed at the cell surface in an enzymically active form, was not shed, implying that the juxtamembrane stalk is the critical determinant of shedding. However, an additional construct, ACEDeltaC, in which the N-terminal domain of somatic ACE was fused to the stalk, TM and cytosolic domains, was also not shed, despite the presence of a cleavable stalk, implying that in contrast with the C-terminal domain, the N-terminal domain lacks a signal required for shedding. These data are discussed in the context of two classes of secretases that differ in their requirements for recognition of substrate proteins." @default.
• W1966307736 created "2016-06-24" @default.
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• W1966307736 date "2001-08-15" @default.
• W1966307736 modified "2023-09-26" @default.
• W1966307736 title "Roles of the juxtamembrane and extracellular domains of angiotensin-converting enzyme in ectodomain shedding" @default.
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• W1966307736 doi "https://doi.org/10.1042/0264-6021:3580185" @default.
• W1966307736 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1222046" @default.
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• W1966307736 hasPublicationYear "2001" @default.
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