A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase

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dc.contributor.authorAlfalah, Marwan
dc.contributor.authorParkin, Edward T
dc.contributor.authorJacob, Ralf
dc.contributor.authorSturrock, Edward D
dc.contributor.authorMentele, Reinhard
dc.contributor.authorTurner, Anthony J
dc.contributor.authorHOOPER, Nigel M
dc.contributor.authorNaim, Hassan Y
dc.date.accessioned2021-10-08T07:20:46Z
dc.date.available2021-10-08T07:20:46Z
dc.date.issued2001
dc.description.abstractAngiotensin I-converting enzyme (ACE) is one of a number of integral membrane proteins that is proteolytically shed from the cell surface by a zinc metallosecretase. Mutagenesis of Asn(631) to Gln in the juxtamembrane stalk region of ACE resulted in more efficient secretion of the mutant protein (ACE(NQ)) as determined by pulse-chase analysis. In contrast to the wild-type ACE, the cleavage of ACE(NQ) was not blocked by the metallosecretase inhibitor batimastat but by the serine protease inhibitor, 1,3-dichloroisocoumarin. Incubation of the cells at 15 degrees C revealed that ACE(NQ) was cleaved in the endoplasmic reticulum, and mass spectrometric analysis of the secreted form of the protein indicated that it had been cleaved at the Asn(635)-Ser(636) bond, three residues N-terminal to the normal secretase cleavage site at Arg(638)-Ser(639). These data clearly show that a point mutation in the juxtamembrane region of an integral membrane protein can invoke the action of a mechanistically and spatially distinct secretase. In light of this observation, previous data on the effect of mutations in the juxtamembrane stalk of shed proteins being accommodated by a single secretase having a relaxed specificity need to be re-evaluated.
dc.identifier.apacitationAlfalah, M., Parkin, E. T., Jacob, R., Sturrock, E. D., Mentele, R., Turner, A. J., ... Naim, H. Y. (2001). A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase. <i>The Journal of Biological Chemistry</i>, 276(24), 21105 - 21109. http://hdl.handle.net/11427/35001en_ZA
dc.identifier.chicagocitationAlfalah, Marwan, Edward T Parkin, Ralf Jacob, Edward D Sturrock, Reinhard Mentele, Anthony J Turner, Nigel M HOOPER, and Hassan Y Naim "A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase." <i>The Journal of Biological Chemistry</i> 276, 24. (2001): 21105 - 21109. http://hdl.handle.net/11427/35001en_ZA
dc.identifier.citationAlfalah, M., Parkin, E.T., Jacob, R., Sturrock, E.D., Mentele, R., Turner, A.J., HOOPER, N.M. & Naim, H.Y. et al. 2001. A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase. <i>The Journal of Biological Chemistry.</i> 276(24):21105 - 21109. http://hdl.handle.net/11427/35001en_ZA
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.ris TY - Journal Article AU - Alfalah, Marwan AU - Parkin, Edward T AU - Jacob, Ralf AU - Sturrock, Edward D AU - Mentele, Reinhard AU - Turner, Anthony J AU - HOOPER, Nigel M AU - Naim, Hassan Y AB - Angiotensin I-converting enzyme (ACE) is one of a number of integral membrane proteins that is proteolytically shed from the cell surface by a zinc metallosecretase. Mutagenesis of Asn(631) to Gln in the juxtamembrane stalk region of ACE resulted in more efficient secretion of the mutant protein (ACE(NQ)) as determined by pulse-chase analysis. In contrast to the wild-type ACE, the cleavage of ACE(NQ) was not blocked by the metallosecretase inhibitor batimastat but by the serine protease inhibitor, 1,3-dichloroisocoumarin. Incubation of the cells at 15 degrees C revealed that ACE(NQ) was cleaved in the endoplasmic reticulum, and mass spectrometric analysis of the secreted form of the protein indicated that it had been cleaved at the Asn(635)-Ser(636) bond, three residues N-terminal to the normal secretase cleavage site at Arg(638)-Ser(639). These data clearly show that a point mutation in the juxtamembrane region of an integral membrane protein can invoke the action of a mechanistically and spatially distinct secretase. In light of this observation, previous data on the effect of mutations in the juxtamembrane stalk of shed proteins being accommodated by a single secretase having a relaxed specificity need to be re-evaluated. DA - 2001 DB - OpenUCT DP - University of Cape Town IS - 24 J1 - The Journal of Biological Chemistry LK - https://open.uct.ac.za PY - 2001 SM - 0021-9258 SM - 1083-351X T1 - A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase TI - A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase UR - http://hdl.handle.net/11427/35001 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/35001
dc.identifier.vancouvercitationAlfalah M, Parkin ET, Jacob R, Sturrock ED, Mentele R, Turner AJ, et al. A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase. The Journal of Biological Chemistry. 2001;276(24):21105 - 21109. http://hdl.handle.net/11427/35001.en_ZA
dc.language.isoeng
dc.publisher.departmentDivision of Medical Biochemistry
dc.publisher.facultyFaculty of Health Sciences
dc.sourceThe Journal of Biological Chemistry
dc.source.journalissue24
dc.source.journalvolume276
dc.source.pagination21105 - 21109
dc.source.urihttps://dx.doi.org/10.1074/jbc.M100339200
dc.subject.otherAmino Acid Sequence
dc.subject.otherAmino Acid Substitution
dc.subject.otherAmyloid Precursor Protein Secretases
dc.subject.otherAspartic Acid Endopeptidases
dc.subject.otherCell Line
dc.subject.otherCell Membrane
dc.subject.otherEndopeptidases
dc.subject.otherHumans
dc.subject.otherKinetics
dc.subject.otherMolecular Sequence Data
dc.subject.otherMutagenesis, Site-Directed
dc.subject.otherNeuroblastoma
dc.subject.otherNeurons
dc.subject.otherPeptidyl-Dipeptidase A
dc.subject.otherPhenylalanine
dc.subject.otherPoint Mutation
dc.titleA Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase
dc.typeJournal Article
uct.type.publicationResearch
uct.type.resourceJournal Article
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