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Dive into the research topics where Sylva L. U. Schwager is active.

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Featured researches published by Sylva L. U. Schwager.


Nature | 2003

Crystal Structure of the Human Angiotensin-Converting Enzyme-Lisinopril Complex

Ramanathan Natesh; Sylva L. U. Schwager; Edward D. Sturrock; K.R. Acharya

Angiotensin-converting enzyme (ACE) has a critical role in cardiovascular function by cleaving the carboxy terminal His-Leu dipeptide from angiotensin I to produce a potent vasopressor octapeptide, angiotensin II. Inhibitors of ACE are a first line of therapy for hypertension, heart failure, myocardial infarction and diabetic nephropathy. Notably, these inhibitors were developed without knowledge of the structure of human ACE, but were instead designed on the basis of an assumed mechanistic homology with carboxypeptidase A. Here we present the X-ray structure of human testicular ACE and its complex with one of the most widely used inhibitors, lisinopril (N2-[(S)-1-carboxy-3-phenylpropyl]-l-lysyl-l-proline; also known as Prinivil or Zestril), at 2.0 Å resolution. Analysis of the three-dimensional structure of ACE shows that it bears little similarity to that of carboxypeptidase A, but instead resembles neurolysin and Pyrococcus furiosus carboxypeptidase—zinc metallopeptidases with no detectable sequence similarity to ACE. The structure provides an opportunity to design domain-selective ACE inhibitors that may exhibit new pharmacological profiles.


Scientific Reports | 2012

Molecular Recognition and Regulation of Human Angiotensin-I Converting Enzyme (Ace) Activity by Natural Inhibitory Peptides.

Geoffrey Masuyer; Sylva L. U. Schwager; Edward D. Sturrock; R. Elwyn Isaac; K. Ravi Acharya

Angiotensin-I converting enzyme (ACE), a two-domain dipeptidylcarboxypeptidase, is a key regulator of blood pressure as a result of its critical role in the renin-angiotensin-aldosterone and kallikrein-kinin systems. Hence it is an important drug target in the treatment of cardiovascular diseases. ACE is primarily known for its ability to cleave angiotensin I (Ang I) to the vasoactive octapeptide angiotensin II (Ang II), but is also able to cleave a number of other substrates including the vasodilator bradykinin and N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP), a physiological modulator of hematopoiesis. For the first time we provide a detailed biochemical and structural basis for the domain selectivity of the natural peptide inhibitors of ACE, bradykinin potentiating peptide b and Ang II. Moreover, Ang II showed selective competitive inhibition of the carboxy-terminal domain of human somatic ACE providing evidence for a regulatory role in the human renin-angiotensin system (RAS).


Biochemical Journal | 2003

Deglycosylation, processing and crystallization of human testis angiotensin-converting enzyme.

Kerry Gordon; Pierre Redelinghuys; Sylva L. U. Schwager; Mario R. W. Ehlers; Anastassios C. Papageorgiou; Ramanathan Natesh; K. Ravi Acharya; Edward D. Sturrock

Angiotensin I-converting enzyme (ACE) is a highly glycosylated type I integral membrane protein. A series of underglycosylated testicular ACE (tACE) glycoforms, lacking between one and five N-linked glycosylation sites, were used to assess the role of glycosylation in tACE processing, crystallization and enzyme activity. Whereas underglycosylated glycoforms showed differences in expression and processing, their kinetic parameters were similar to that of native tACE. N-glycosylation of Asn-72 or Asn-109 was necessary and sufficient for the production of enzymically active tACE but glycosylation of Asn-90 alone resulted in rapid intracellular degradation. All mutants showed similar levels of phorbol ester stimulation and were solubilized at the same juxtamembrane cleavage site as the native enzyme. Two mutants, tACEDelta36-g1234 and -g13, were successfully crystallized, diffracting to 2.8 and 3.0 A resolution respectively. Furthermore, a truncated, soluble tACE (tACEDelta36NJ), expressed in the presence of the glucosidase-I inhibitor N -butyldeoxynojirimycin, retained the activity of the native enzyme and yielded crystals belonging to the orthorhombic P2(1)2(1)2(1) space group (cell dimensions, a=56.47 A, b=84.90 A, c=133.99 A, alpha=90 degrees, beta=90 degrees and gamma=90 degrees ). These crystals diffracted to 2.0 A resolution. Thus underglycosylated human tACE mutants, lacking O-linked oligosaccharides and most N-linked oligosaccharides or with only simple N-linked oligosaccharides attached throughout the molecule, are suitable for X-ray diffraction studies.


Journal of Biological Chemistry | 2010

The N Domain of Human Angiotensin-I-converting Enzyme: THE ROLE OF N-GLYCOSYLATION AND THE CRYSTAL STRUCTURE IN COMPLEX WITH AN N DOMAIN-SPECIFIC PHOSPHINIC INHIBITOR, RXP407*

Colin S. Anthony; Hazel R. Corradi; Sylva L. U. Schwager; Pierre Redelinghuys; Dimitris Georgiadis; Vincent Dive; K. Ravi Acharya; Edward D. Sturrock

Angiotensin-I-converting enzyme (ACE) plays a critical role in the regulation of blood pressure through its central role in the renin-angiotensin and kallikrein-kinin systems. ACE contains two domains, the N and C domains, both of which are heavily glycosylated. Structural studies of ACE have been fraught with severe difficulties because of surface glycosylation of the protein. In order to investigate the role of glycosylation in the N domain and to create suitable forms for crystallization, we have investigated the importance of the 10 potential N-linked glycan sites using enzymatic deglycosylation, limited proteolysis, and mass spectrometry. A number of glycosylation mutants were generated via site-directed mutagenesis, expressed in CHO cells, and analyzed for enzymatic activity and thermal stability. At least eight of 10 of the potential glycan sites are glycosylated; three C-terminal sites were sufficient for expression of active N domain, whereas two N-terminal sites are important for its thermal stability. The minimally glycosylated Ndom389 construct was highly suitable for crystallization studies. The structure in the presence of an N domain-selective phosphinic inhibitor RXP407 was determined to 2.0 Å resolution. The Ndom389 structure revealed a hinge region that may contribute to the breathing motion proposed for substrate binding.


Biochemical Journal | 2001

Roles of the juxtamembrane and extracellular domains of angiotensin-converting enzyme in ectodomain shedding

Susan Pang; Anthony J. Chubb; Sylva L. U. Schwager; Mario R. W. Ehlers; Edward D. Sturrock; Nigel M. Hooper

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.


FEBS Journal | 2011

Structural Characterization of Angiotensin-I Converting Enzyme in Complex with a Selenium Analogue of Captopril

Mohd Akif; Geoffrey Masuyer; Sylva L. U. Schwager; Bhaskar J. Bhuyan; Govindasamy Mugesh; R. Elwyn Isaac; Edward D. Sturrock; K. Ravi Acharya

Human somatic angiotensin I‐converting enzyme (ACE), a zinc‐dependent dipeptidyl carboxypeptidase, is central to the regulation of the renin–angiotensin aldosterone system. It is a well‐known target for combating hypertension and related cardiovascular diseases. In a recent study by Bhuyan and Mugesh [Org. Biomol. Chem. (2011) 9, 1356–1365], it was shown that the selenium analogues of captopril (a well‐known clinical inhibitor of ACE) not only inhibit ACE, but also protect against peroxynitrite‐mediated nitration of peptides and proteins. Here, we report the crystal structures of human testis ACE (tACE) and a homologue of ACE, known as AnCE, from Drosophila melanogaster in complex with the most promising selenium analogue of captopril (SeCap) determined at 2.4 and 2.35 Å resolution, respectively. The inhibitor binds at the active site of tACE and AnCE in an analogous fashion to that observed for captopril and provide the first examples of a protein–selenolate interaction. These new structures of tACE–SeCap and AnCE–SeCap inhibitor complexes presented here provide important information for further exploration of zinc coordinating selenium‐based ACE inhibitor pharmacophores with significant antioxidant activity.


Biochemical and Biophysical Research Communications | 2002

Defining the boundaries of the testis angiotensin I-converting enzyme ectodomain

Anthony J. Chubb; Sylva L. U. Schwager; Zenda L. Woodman; Mario R. W. Ehlers; Edward D. Sturrock

Numerous cytokines, receptors, and ectoenzymes, including angiotensin I-converting enzyme (ACE), are shed from the cell surface by limited proteolysis at the juxtamembrane stalk region. The membrane-proximal C domain of ACE has been implicated in sheddase-substrate recognition. We mapped the functional boundaries of the testis ACE ectodomain (identical to the C domain of somatic ACE) by progressive deletions from the N- and C-termini and analysing the effects on catalytic activity, stability, and shedding in transfected cells. We found that deletions extending beyond Leu37 at the N-terminus and Trp616 at the C-terminus abolished catalytic activity and shedding, either by disturbing the ectodomain conformation or by inhibiting maturation and surface expression. Based on these data and on sequence alignments, we propose that the boundaries of the ACE ectodomain are Asp40 at the N-terminus and Gly615 at the C-terminus.


Biochimica et Biophysica Acta | 1983

The isolation of isohistones by preparative gel electrophoresis from embryos of the sea urchin Parechinus angulosus

Sylva L. U. Schwager; Wolf F. Brandt; Claus von Holt

Abstract Ten isohistones from the embryo of the sea urchin Parechinus angulosus have been isolated by preparative gel electrophoresis and characterised by electrophoretic mobility in two detergent systems, amino acid composition and partial sequences. This brings the total number of different histones identified which are synthesized at one or the other time during the life cycle of the sea urchin to a minimum of 24 structurally characterised polypeptides.


Journal of Clinical Hypertension | 2013

Association of B2 receptor polymorphisms and ACE activity with ACE inhibitor-induced angioedema in black and mixed-race South Africans.

Retsilisitsoe R. Moholisa; Brian R. Rayner; E. Patricia Owen; Sylva L. U. Schwager; Joalice S. Stark; Motassim Badri; Clint L. Cupido; Edward D. Sturrock

Angiotensin‐converting enzyme (ACE) inhibitors are first‐line therapy for the treatment of hypertension, congestive heart failure, and diabetic nephropathy. ACE inhibitors are associated with adverse side effects such as persistent dry cough (ACE‐cough) and, rarely, life‐threatening angioedema (ACE‐AE). The authors investigated the influence of ACE I/D polymorphism in combination with serum ACE activity, B2 receptor −9/+9 polymorphism, and B2 receptor C‐58T single nucleotide polymorphism (SNP) on the development of ACE‐AE and ACE‐cough. The frequencies of ACE I/D as well as B2 receptor +9/−9 and C‐58T polymorphisms were compared in patients with ACE‐AE, ACE‐cough, and ACE inhibitor–exposed controls, and serum ACE activity was measured. There were 52 cases of ACE‐AE, 36 cases of ACE‐cough, and 77 controls. The genotyping revealed a significant association between the B2 −9 allele and ACE inhibitor–induced AE (62% vs 38%, P=.008), and ACE inhibitor–induced cough (61% vs 38%, P=.02) when compared with controls. There was no significant association between ACE I/D polymorphism as well as the B2 C‐58T SNP with both ACE‐induced AE and cough. ACE activity was significantly higher in controls compared with patients with ACE‐AE (34.5±1.14 mU/mL vs 17.8±0.86 mU/mL, P=.0001) and ACE‐cough (34.5±1.14 mU/mL vs 23.3±1.88 mU/mL, P=.0001). Thus, our data suggest that the B2 −9 allele and reduced ACE activity are associated with both ACE‐AE and ACE‐cough.


Biological Chemistry | 2006

Homologous substitution of ACE C-domain regions with N-domain sequences: effect on processing, shedding, and catalytic properties

Zenda L. Woodman; Sylva L. U. Schwager; Pierre Redelinghuys; Anthony J. Chubb; Elizabeth L. van der Merwe; Mario R. W. Ehlers; Edward D. Sturrock

Abstract Angiotensin-converting enzyme (ACE) exists as two isoforms: somatic ACE (sACE), comprised of two homologous N and C domains, and testis ACE (tACE), comprised of the C domain only. The N and C domains are both active, but show differences in substrate and inhibitor specificity. While both isoforms are shed from the cell surface via a sheddase-mediated cleavage, tACE is shed much more efficiently than sACE. To delineate the regions of tACE that are important in catalytic activity, intracellular processing, and regulated ectodomain shedding, regions of the tACE sequence were replaced with the corresponding N-domain sequence. The resultant chimeras C1–163Ndom-ACE, C417–579Ndom-ACE, and C583–623Ndom-ACE were processed to the cell surface of transfected Chinese hamster ovary (CHO) cells, and were cleaved at the identical site as that of tACE. They also showed acquisition of N-domain-like catalytic properties. Homology modelling of the chimeric proteins revealed structural changes in regions required for tACE-specific catalytic activity. In contrast, C164–416Ndom-ACE and C191–214Ndom-ACE demonstrated defective intracellular processing and were neither enzymatically active nor shed. Therefore, critical elements within region D164–V416 and more specifically I191–T214 are required for the processing, cell-surface targeting, and enzyme activity of tACE, and cannot be substituted for by the homologous N-domain sequence.

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Anthony J. Chubb

Royal College of Surgeons in Ireland

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