Structure of human neutral endopeptidase (neprilysin) complexed with phosphoramidon 1 1Edited by R. Huber

Oefner, Christian; D’Arcy, Allan; Hennig, Michael; Winkler, Fritz K.; Dale, Glenn E.

doi:10.1006/jmbi.1999.3492

Cited by 262 publications

(288 citation statements)

References 48 publications

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“…The three-dimensional (3D) structure of a part of PHEX based on the published 3D structure of NEP1 (PBD 1DMT). 26 Highlighted in red are residues of the active site. Pro534 is indicated in blue.…”

Section: Discussionmentioning

confidence: 99%

“…25 Additionally, the molecular visualisation software PyMol 0.99rc6 (www.pymol.org) was used to predict the functional consequence of the missense mutation on the tertiary structure at the site of the mutation. The 3D structure of NEP was described by Oefner et al 26 Mutations fulfilling criteria two and/or three only were considered novel.…”

Section: Genetic Analysismentioning

confidence: 99%

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Mutational analysis of PHEX, FGF23, DMP1, SLC34A3 and CLCN5 in patients with hypophosphatemic rickets

et al. 2012

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This study aimed to identify the underlying genetic mutation in patients with hypophosphatemic rickets (HR). Genomic DNA was analysed for mutations in PHEX, FGF23 and CLCN5 by polymerase chain reaction (PCR) followed by denaturing highperformance liquid chromatography (dHPLC). Bi-directional sequencing was performed in samples with deviating chromatographic profiles. DMP1 and SLC34A3 were sequenced, only. In addition, a multiplex ligation-dependent probe amplification (MLPA) analysis was performed to detect larger deletions/duplications in PHEX or FGF23. Familial cases accounted for 12 probands while 12 cases were sporadic. In 20 probands, mutations were detected in PHEX of which 12 were novel, and one novel frameshift mutation was found in DMP1. Three PHEX mutations were identified by the MLPA analysis only; that is, two large deletions and one duplication. No mutations were identified in FGF23, SLC34A3 or CLCN5. By the methods used, a disease causing mutation was identified in 83% of the familial and 92% of the sporadic cases, thereby in 88% of the tested probands. Genetic analysis performed in HR patients by PCR, dHPLC, sequencing and in addition by MLPA analysis revealed a high identification rate of gene mutations causing HR, including 12 novel PHEX and one novel DMP1 mutation.

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Section: Discussionmentioning

confidence: 99%

Section: Genetic Analysismentioning

confidence: 99%

Mutational analysis of PHEX, FGF23, DMP1, SLC34A3 and CLCN5 in patients with hypophosphatemic rickets

et al. 2012

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“…The hydrogen bond between the side chain of the acidic glutamate and the protonated nitrogen HN ␦1 atom of the imidazole ring of His-385 (the equivalent of His-295 in LTA 4 H) could function both by maintaining the position of the histidine side chain relative to the zinc ion and by polarizing the histidine N ␦2 atom, thereby increasing the strength of zinc coordination (36). This suggests that Glu-415 of APA may be functionally equivalent to Asp-991 in angiotensin-converting enzyme, Asp-170 in thermolysin, and Asp-650 in neutral endopeptidase 24.11, all of which are located in the conserved motif EXXXD and have been shown to play a role in positioning the first histidine (His-959 in angiotensin-converting enzyme, His-142 in thermolysin, and His-583 in neutral endopeptidase 24.11) of their respective zinc-binding motif HEXXH (26,37,38). In addition, the distance between the third zinc ligand and this acidic residue is conserved among the gluzincin family (four residues in angiotensin-converting enzyme, thermolysin, and neutral endopeptidase 24.11 and seven residues in APA, LTA 4 H, and other monozinc aminopeptidases), suggesting a common functional role for this residue in monozinc aminopeptidases.…”

Section: Discussionmentioning

confidence: 99%

Contribution of Molecular Modeling and Site-directed Mutagenesis to the Identification of Two Structural Residues, Arg-220 and Asp-227, in Aminopeptidase A

Rozenfeld¹,

Iturrioz²,

Maigret³

et al. 2002

Journal of Biological Chemistry

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Aminopeptidase A is a zinc metalloenzyme involved in the formation of brain angiotensin III, which exerts a tonic stimulatory action on the central control of blood pressure. Thus, central inhibitors of aminopeptidase A constitute putative central antihypertensive agents. Mutagenic studies have been performed to investigate organization of the aminopeptidase A active site, with a view to designing such inhibitors. The structure of one monozinc aminopeptidase (leukotriene A 4 hydrolase) was recently resolved and used to construct a threedimensional model of the aminopeptidase A ectodomain. This new model, highly consistent with the results of mutagenic studies, showed a critical structural interaction between two conserved residues, Arg-220 and Asp-227. Mutagenic replacement of either of these two residues disrupted maturation and subcellular localization and abolished the enzymatic activity of aminopeptidase A, confirming the critical structural role of these residues. In this study, we generated the first threedimensional model of a strict aminopeptidase, aminopeptidase A. This model constitutes a new tool to probe further the active site of aminopeptidase A and to design new inhibitors of this enzyme.

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“…(1)(2)(3) In humans, inactivating mutations in PHEX cause X-linked hypophosphatemia (XLH), with similar phenotypic features occurring in the hypophosphatemic (Hyp) mouse model of this disease. XLH is the most frequent inherited formin mineral ion metabolism and local changes within the bone tissue, resulting in the clinical and biochemical features of XLH, are still not completely known.…”

Section: Introductionmentioning

confidence: 99%

Proteolytic processing of osteopontin by PHEX and accumulation of osteopontin fragments in Hyp mouse bone, the murine model of X-linked hypophosphatemia

Barros

Hoac

Neves

et al. 2012

Journal of Bone and Mineral Research

125

111

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X-linked hypophosphatemia (XLH/HYP)-with renal phosphate wasting, hypophosphatemia, osteomalacia, and tooth abscesses-is caused by mutations in the zinc-metallopeptidase PHEX gene (phosphate-regulating gene with homologies to endopeptidase on the X chromosome). PHEX is highly expressed by mineralized tissue cells. Inactivating mutations in PHEX lead to distal renal effects (implying accumulation of a secreted, circulating phosphaturic factor) and accumulation in bone and teeth of mineralization-inhibiting, acidic serine-and aspartate-rich motif (ASARM)-containing peptides, which are proteolytically derived from the mineral-binding matrix proteins of the SIBLING family (small, integrin-binding ligand N-linked glycoproteins). Although the latter observation suggests a local, direct matrix effect for PHEX, its physiologically relevant substrate protein(s) have not been identified. Here, we investigated two SIBLING proteins containing the ASARM motif-osteopontin (OPN) and bone sialoprotein (BSP)-as potential substrates for PHEX. Using cleavage assays, gel electrophoresis, and mass spectrometry, we report that OPN is a full-length protein substrate for PHEX. Degradation of OPN was essentially complete, including hydrolysis of the ASARM motif, resulting in only very small residual fragments. Western blotting of Hyp (the murine homolog of human XLH) mouse bone extracts having no PHEX activity clearly showed accumulation of an $35 kDa OPN fragment that was not present in wild-type mouse bone. Immunohistochemistry and immunogold labeling (electron microscopy) for OPN in Hyp bone likewise showed an accumulation of OPN and/or its fragments compared with normal wild-type bone. Incubation of Hyp mouse bone extracts with PHEX resulted in the complete degradation of these fragments. In conclusion, these results identify full-length OPN and its fragments as novel, physiologically relevant substrates for PHEX, suggesting that accumulation of mineralization-inhibiting OPN fragments may contribute to the mineralization defect seen in the osteomalacic bone characteristic of XLH/HYP. ß

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Structure of human neutral endopeptidase (neprilysin) complexed with phosphoramidon 1 1Edited by R. Huber

Cited by 262 publications

References 48 publications

Mutational analysis of PHEX, FGF23, DMP1, SLC34A3 and CLCN5 in patients with hypophosphatemic rickets

Mutational analysis of PHEX, FGF23, DMP1, SLC34A3 and CLCN5 in patients with hypophosphatemic rickets

Contribution of Molecular Modeling and Site-directed Mutagenesis to the Identification of Two Structural Residues, Arg-220 and Asp-227, in Aminopeptidase A

Proteolytic processing of osteopontin by PHEX and accumulation of osteopontin fragments in Hyp mouse bone, the murine model of X-linked hypophosphatemia

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