A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP

Simple item page
dc.contributor.authorDanilov, Sergei Men_ZA
dc.contributor.authorWade, Michael Sen_ZA
dc.contributor.authorSchwager, Sylva Len_ZA
dc.contributor.authorDouglas, Ross Gen_ZA
dc.contributor.authorNesterovitch, Andrew Ben_ZA
dc.contributor.authorPopova, Isolda Aen_ZA
dc.contributor.authorHogarth, Kyle Den_ZA
dc.contributor.authorBhardwaj, Nakulen_ZA
dc.contributor.authorSchwartz, David Een_ZA
dc.contributor.authorSturrock, Edward Den_ZA
dc.contributor.authorGarcia, Joe G Nen_ZA
dc.date.accessioned2016-01-11T06:53:37Z
dc.date.available2016-01-11T06:53:37Z
dc.date.issued2014en_ZA
dc.description.abstractBACKGROUND: Angiotensin I-converting enzyme (ACE) has two functional N- and C-domain active centers that display differences in the metabolism of biologically-active peptides including the hemoregulatory tetrapeptide, Ac-SDKP, hydrolysed preferentially by the N domain active center. Elevated Ac-SDKP concentrations are associated with reduced tissue fibrosis. RESULTS: We identified a patient of African descent exhibiting unusual blood ACE kinetics with reduced relative hydrolysis of two synthetic ACE substrates (ZPHL/HHL ratio) suggestive of the ACE N domain center inactivation. Inhibition of blood ACE activity by anti-catalytic mAbs and ACE inhibitors and conformational fingerprint of blood ACE suggested overall conformational changes in the ACE molecule and sequencing identified Ser333Trp substitution in the N domain of ACE. In silico analysis demonstrated S333W localized in the S 1 pocket of the active site of the N domain with the bulky Trp adversely affecting binding of ACE substrates due to steric hindrance. Expression of mutant ACE (S333W) in CHO cells confirmed altered kinetic properties of mutant ACE and conformational changes in the N domain. Further, the S333W mutant displayed decreased ability (5-fold) to cleave the physiological substrate AcSDKP compared to wild-type ACE. Conclusions and Significance A novel Ser333Trp ACE mutation results in dramatic changes in ACE kinetic properties and lowered clearance of Ac-SDKP. Individuals with this mutation (likely with significantly increased levels of the hemoregulatory tetrapeptide in blood and tissues), may confer protection against fibrosis.en_ZA
dc.identifier.apacitationDanilov, S. M., Wade, M. S., Schwager, S. L., Douglas, R. G., Nesterovitch, A. B., Popova, I. A., ... Garcia, J. G. N. (2014). A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP. <i>PLoS One</i>, http://hdl.handle.net/11427/16286en_ZA
dc.identifier.chicagocitationDanilov, Sergei M, Michael S Wade, Sylva L Schwager, Ross G Douglas, Andrew B Nesterovitch, Isolda A Popova, Kyle D Hogarth, et al "A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP." <i>PLoS One</i> (2014) http://hdl.handle.net/11427/16286en_ZA
dc.identifier.citationDanilov, S. M., Wade, M. S., Schwager, S. L., Douglas, R. G., Nesterovitch, A. B., Popova, I. A., ... & Garcia, J. G. (2014). A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP. PloS one, 9(2), e88001. doi:10.1371/journal.pone.0088001en_ZA
dc.identifier.ris TY - Journal Article AU - Danilov, Sergei M AU - Wade, Michael S AU - Schwager, Sylva L AU - Douglas, Ross G AU - Nesterovitch, Andrew B AU - Popova, Isolda A AU - Hogarth, Kyle D AU - Bhardwaj, Nakul AU - Schwartz, David E AU - Sturrock, Edward D AU - Garcia, Joe G N AB - BACKGROUND: Angiotensin I-converting enzyme (ACE) has two functional N- and C-domain active centers that display differences in the metabolism of biologically-active peptides including the hemoregulatory tetrapeptide, Ac-SDKP, hydrolysed preferentially by the N domain active center. Elevated Ac-SDKP concentrations are associated with reduced tissue fibrosis. RESULTS: We identified a patient of African descent exhibiting unusual blood ACE kinetics with reduced relative hydrolysis of two synthetic ACE substrates (ZPHL/HHL ratio) suggestive of the ACE N domain center inactivation. Inhibition of blood ACE activity by anti-catalytic mAbs and ACE inhibitors and conformational fingerprint of blood ACE suggested overall conformational changes in the ACE molecule and sequencing identified Ser333Trp substitution in the N domain of ACE. In silico analysis demonstrated S333W localized in the S 1 pocket of the active site of the N domain with the bulky Trp adversely affecting binding of ACE substrates due to steric hindrance. Expression of mutant ACE (S333W) in CHO cells confirmed altered kinetic properties of mutant ACE and conformational changes in the N domain. Further, the S333W mutant displayed decreased ability (5-fold) to cleave the physiological substrate AcSDKP compared to wild-type ACE. Conclusions and Significance A novel Ser333Trp ACE mutation results in dramatic changes in ACE kinetic properties and lowered clearance of Ac-SDKP. Individuals with this mutation (likely with significantly increased levels of the hemoregulatory tetrapeptide in blood and tissues), may confer protection against fibrosis. DA - 2014 DB - OpenUCT DO - 10.1371/journal.pone.0088001 DP - University of Cape Town J1 - PLoS One LK - https://open.uct.ac.za PB - University of Cape Town PY - 2014 T1 - A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP TI - A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP UR - http://hdl.handle.net/11427/16286 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/16286
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0088001
dc.identifier.vancouvercitationDanilov SM, Wade MS, Schwager SL, Douglas RG, Nesterovitch AB, Popova IA, et al. A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP. PLoS One. 2014; http://hdl.handle.net/11427/16286.en_ZA
dc.language.isoengen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.publisher.departmentInstitute of Infectious Disease and Molecular Medicineen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_ZA
dc.rights.holder© 2014 Danilov et alen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0en_ZA
dc.sourcePLoS Oneen_ZA
dc.source.urihttp://journals.plos.org/plosoneen_ZA
dc.subject.otherBlooden_ZA
dc.subject.otherACE inhibitorsen_ZA
dc.subject.otherHydrolysisen_ZA
dc.subject.otherSubstitution mutationen_ZA
dc.subject.otherCHO cellsen_ZA
dc.subject.otherMembrane proteinsen_ZA
dc.subject.otherMutation detectionen_ZA
dc.subject.otherCrystal structureen_ZA
dc.titleA novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKPen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Danilov_A_Novel_Angiotensin_I_2014.pdf
Size:
1.38 MB
Format:
Adobe Portable Document Format
Description:
Download
Collections
Journal Articles