Rat insulin-degrading enzyme: cleavage pattern of the natriuretic peptide hormones ANP, BNP, and CNP revealed by HPLC and mass spectrometry

Müller, Dieter; Schulze, Christian; Baumeister, Hans; Buck, Fritz; Richter, Dietmar

doi:10.1021/bi00160a026

Cited by 71 publications

(58 citation statements)

References 35 publications

(43 reference statements)

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“…Endopeptidase mRNA is present at early stages of spermatogenesis, whereas no enzyme immunoreactivity is observed in spermatogonia and spermatocytes (13 (34,35,38), NRD convertase is also distantly related to MPPs, which have been implicated in mitochondrial processing (39). We therefore suggest that the NRD convertase may constitute the prototype of a distinct class of processing enzymes.…”

Section: Discussionmentioning

confidence: 86%

“…The presence in the deduced amino acid sequence of the zinc-binding motif confirms the metalloenzyme character of NRD convertase. In addition to their sequence homology, IDEs and NRD convertase share several biochemical characteristics such as their capacity to cleave peptide bonds on the N terminus of a limited number of residues and their sensitivity to some sulfhydryl reagents (34). The alignment of NRD convertase and the three IDE sequences identified a unique Cys residue (Cys-959, Fig.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

N-arginine dibasic convertase, a metalloendopeptidase as a prototype of a class of processing enzymes.

Pierotti

Prat

Chesneau

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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N-Arg dibasic convertase is a me tidase from rat brain cortex and testis that cleaves peptide substrates on the N terminu ofArg residues in dibasic s s.By using both an oinu ide and antibodies to screen a rat testis cDNA library, a fill-length cDNA (4,5). On the basis of its similarity to subtilisin and to furin (6), a human homolog of the Kex2 protein, a family of prohormone convertases (PCs) has been identified by PCR techniques. Their involvement in processing of a number of propeptides and proproteins was inferred mainly from cotransfection experiments (7-9), and for PC1, by the use of antisense mRNA (10).Characterization of putative processing endoproteases by classical biochemical techniques has led to the identification of a number of activities, selective for basic residues in precursors, that belong to the four classes of proteases (metallo-, serine, aspartyl, and thiol enzymes; for review, see ref. 11). This suggested that more than one processing endoprotease family could exist (12). To our knowledge, none of these basic-residue-specific enzymes has been cloned.Recently, a metalloendopeptidase was completely purified from rat testis and shown to cleave a number of peptide substrates on the N terminus of Arg residues in dibasic moieties (13). This enzyme was also present in rat brain cortex and its functional properties appeared undistinguishable from those of the somatostatin-28 convertase activity previously identified in this tissue (14,15). By using microsequencing of tryptic fragments of the purified enzyme to design an oligonucleotide probe and polyclonal antibodies raised against the purified protein (13) (20)].The in vitro highly restricted specificity ofNRD convertase for Arg residues in dibasic processing signals and its belonging to the M16 family, which contains other enzymes involved in maturation, suggest that this enzyme is the prototype of a distinct family of processing endoproteases. MATERIALS AND METHODSIsolatin and Cherizatin of cDNA Cones E d NRD Convertae. Four tryptic fragments were sequenced after digestion of previously purified NRD convertase following native PAGE. One fragment, GMQLIYLPPSPLLAE, was used to design the following degenerate inosine-containing oligonucleotide: 5'-GGIGGIAG(A/G)TAIATIA(G/A)(T/ C)TGCATICC-3'. Two additional peptides were obtained by endolysine C treatment (13) autoradiography of the filters, positive phage plaques were isolated and rescreened to obtain single purified phage isolates. A similar aliquot ofthe library was plated onto NZ-amine medium plates for screening using polyclonal antibodies raised Abbreviations: NRD convertase, N-arginine dibasic convertase; IDE, insulin degrading enzyme; MPP, mitochondrial matrixprocessing peptidase.

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Section: Discussionmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

N-arginine dibasic convertase, a metalloendopeptidase as a prototype of a class of processing enzymes.

Pierotti

Prat

Chesneau

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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“…Comparison of substrate and cleavage-site specificities of pitrilysin with those of insulysin Insulysin (insulin-degrading enzyme, IDE), which is a dimeric or trimeric enzyme, 29) has been reported to degrade not only insulin 30) but also other peptide hormones, such as glucagon, 31,32) ANP, 33,34) transforming growth factor (TGF-), 35) -EP, 36) and calcitonin.…”

Section: Discussionmentioning

confidence: 99%

Cleavage of Various Peptides with Pitrilysin fromEscherichia coli: Kinetic Analyses Using β-Endorphin and Its Derivatives

Cornista¹,

Ikeuchi²,

Haruki³

et al. 2004

Bioscience, Biotechnology, and Biochemistry

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Pitrilysin from Escherichia coli was overproduced, purified, and analyzed for enzymatic activity using 14 peptides as a substrate. Pitrilysin cleaved all the peptides, except for two of the smallest, at a limited number of sites, but showed little amino acid specificity. It cleaved -endorphin (-EP) most effectively, with a K m value of 0.36 M and a k cat value of 750 min À1 . -EP consists of 31 residues and was predominantly cleaved by the enzyme at Lys 19 -Asn 20 . Kinetic analyses using a series of -EP derivatives with N and/or C-terminal truncations and with amino acid substitutions revealed that three hydrophobic residues (Leu 14 , Val 15 , and Leu 17 ) and the region 22-26 in -EP are responsible for high-affinity recognition by the enzyme. These two regions are located on the N-and C-terminal sides of the cleavage site in -EP, suggesting that the substrate binding pocket of pitrilysin spans its catalytic site.

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“…Studies have reported various IDE high-affinity substrates, which were polypeptides 28-51 amino acids in length. A variety of physiological peptides have been shown to be substrates in vitro for IDE including insulin and glucagons (4), the atrial natriuretic factor (5), transforming growth factor · (6,7), the atrial natriuretic peptide (8), ß-endorphin and dynorphins (9), amylin (10), and amyloid ß peptides (11,12). IDE may also have other biological functions such as the clearance of the ß-amyloid precursor protein in the intracellular domain (AICD), which is suggested to regulate the transcriptional activity in the nucleus (13).…”

Section: Introductionmentioning

confidence: 99%

Significant change in insulin production, glucose tolerance and ER stress signaling in transgenic mice coexpressing insulin-siRNA and human IDE

Hwang¹,

Seo

Kim

et al. 2007

Int J Mol Med

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Abstract. The dual expression system for the suppression and clearance of insulin has not been previously used to produce transgenic mice for diabetes-related disease. The aim of this study was to produce new transgenic mice coexpressing specific insulin small interfering RNA (siRNA) sequences and the human insulin degrading enzyme (hIDE) gene in order to examine the diabetes-like phenotype. To achieve this, a new lineage of transgenic mice was produced by the microinjection of the dual expression constructs (pH1/siRNA insulin -CMV/hIDE) into mouse fertilized eggs. The results showed that overexpressing the insulin siRNA and hIDE genes resulted in the induction of the human enzyme, impaired glucose tolerance and lower serum insulin levels compared to the Non-Tg mice. Moreover, the Tg mice aged 20 weeks had a significantly activated ER stress signaling compared to their Non-Tg counterparts, which may be associated with the suppression of insulin production in the pancreas and the degradation of insulin in the liver, respectively. Therefore, insulin-suppressed transgenic mice can be used to examine diabetes as a new diabetes-like phenotype model, which results in a lower level of circulating insulin without the destruction of pancreatic islets.

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Rat insulin-degrading enzyme: cleavage pattern of the natriuretic peptide hormones ANP, BNP, and CNP revealed by HPLC and mass spectrometry

Cited by 71 publications

References 35 publications

N-arginine dibasic convertase, a metalloendopeptidase as a prototype of a class of processing enzymes.

N-arginine dibasic convertase, a metalloendopeptidase as a prototype of a class of processing enzymes.

Cleavage of Various Peptides with Pitrilysin fromEscherichia coli: Kinetic Analyses Using β-Endorphin and Its Derivatives

Significant change in insulin production, glucose tolerance and ER stress signaling in transgenic mice coexpressing insulin-siRNA and human IDE

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