The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis
Insulin-degrading enzyme (IDE) is central to the turnover of insulin and degrades amyloid β (Aβ) in the mammalian brain. Biochemical and genetic data support the notion that IDE may play a role in late onset Alzheimer disease (AD), and recent studies suggest an association between AD and diabetes me...
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todo:paper_00219258_v283_n25_p17039_Llovera2023-10-03T14:23:12Z The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis Llovera, R.E. De Tullio, M. Alonso, L.G. Leissring, M.A. Kaufman, S.B. Roher, A.E. Gay, G.D.P. Morelli, L. Castaño, E.M. Amines Binding energy Binding sites Colloids Diffusers (optical) Enzymes Flow interactions Forming Gelation Glycoproteins High performance liquid chromatography Hormones Insulin Mammals Mass spectrometry A stables After treatments Alzheimer diseases Catalytic domains Catalytic sites Complex formations Cortical tissues Degrading enzymes Diabetes Mellitus Genetic datums Limited proteolyses Mammalian brains Molar excesses Partial denaturations Rat brains Soluble fractions Substrate bindings Terminal fragments Integrodifferential equations amyloid beta protein dodecyl sulfate sodium insulinase recombinant enzyme Alzheimer disease amino terminal sequence article binding site complex formation controlled study enzyme active site enzyme denaturation human human tissue nonhuman pathogenesis priority journal protein conformation protein degradation protein domain protein folding protein protein interaction rat animal biological model brain chemistry enzyme specificity kinetics mass spectrometry metabolism pathology protein binding radiation scattering Mammalia Rattus Alzheimer Disease Amyloid beta-Protein Animals Binding Sites Brain Catalytic Domain Humans Insulysin Kinetics Mass Spectrometry Models, Biological Protein Binding Rats Scattering, Radiation Substrate Specificity Insulin-degrading enzyme (IDE) is central to the turnover of insulin and degrades amyloid β (Aβ) in the mammalian brain. Biochemical and genetic data support the notion that IDE may play a role in late onset Alzheimer disease (AD), and recent studies suggest an association between AD and diabetes mellitus type 2. Here we show that a natively folded recombinant IDE was capable of forming a stable complex with Aβ that resisted dissociation after treatment with strong denaturants. This interaction was also observed with rat brain IDE and detected in an SDS-soluble fraction from AD cortical tissue. Aβ sequence 17-27, known to be crucial in amyloid assembly, was sufficient to form a stable complex with IDE. Monomeric as opposed to aggregated Aβ was competent to associate irreversibly with IDE following a very slow kinetics (t1/2 ∼ 45 min). Partial denaturation of IDE as well as preincubation with a 10-fold molar excess of insulin prevented complex formation, suggesting that the irreversible interaction of Aβ takes place with at least part of the substrate binding site of the protease. Limited proteolysis showed that Aβ remained bound to a ∼25-kDa N-terminal fragment of IDE in an SDS-resistant manner. Mass spectrometry after in gel digestion of the IDE·Aβ complex showed that peptides derived from the region that includes the catalytic site of IDE were recovered with Aβ. Taken together, these results are suggestive of an unprecedented mechanism of conformation-dependent substrate binding that may perturb Aβ clearance, insulin turnover, and promote AD pathogenesis. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00219258_v283_n25_p17039_Llovera |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Amines Binding energy Binding sites Colloids Diffusers (optical) Enzymes Flow interactions Forming Gelation Glycoproteins High performance liquid chromatography Hormones Insulin Mammals Mass spectrometry A stables After treatments Alzheimer diseases Catalytic domains Catalytic sites Complex formations Cortical tissues Degrading enzymes Diabetes Mellitus Genetic datums Limited proteolyses Mammalian brains Molar excesses Partial denaturations Rat brains Soluble fractions Substrate bindings Terminal fragments Integrodifferential equations amyloid beta protein dodecyl sulfate sodium insulinase recombinant enzyme Alzheimer disease amino terminal sequence article binding site complex formation controlled study enzyme active site enzyme denaturation human human tissue nonhuman pathogenesis priority journal protein conformation protein degradation protein domain protein folding protein protein interaction rat animal biological model brain chemistry enzyme specificity kinetics mass spectrometry metabolism pathology protein binding radiation scattering Mammalia Rattus Alzheimer Disease Amyloid beta-Protein Animals Binding Sites Brain Catalytic Domain Humans Insulysin Kinetics Mass Spectrometry Models, Biological Protein Binding Rats Scattering, Radiation Substrate Specificity |
| spellingShingle |
Amines Binding energy Binding sites Colloids Diffusers (optical) Enzymes Flow interactions Forming Gelation Glycoproteins High performance liquid chromatography Hormones Insulin Mammals Mass spectrometry A stables After treatments Alzheimer diseases Catalytic domains Catalytic sites Complex formations Cortical tissues Degrading enzymes Diabetes Mellitus Genetic datums Limited proteolyses Mammalian brains Molar excesses Partial denaturations Rat brains Soluble fractions Substrate bindings Terminal fragments Integrodifferential equations amyloid beta protein dodecyl sulfate sodium insulinase recombinant enzyme Alzheimer disease amino terminal sequence article binding site complex formation controlled study enzyme active site enzyme denaturation human human tissue nonhuman pathogenesis priority journal protein conformation protein degradation protein domain protein folding protein protein interaction rat animal biological model brain chemistry enzyme specificity kinetics mass spectrometry metabolism pathology protein binding radiation scattering Mammalia Rattus Alzheimer Disease Amyloid beta-Protein Animals Binding Sites Brain Catalytic Domain Humans Insulysin Kinetics Mass Spectrometry Models, Biological Protein Binding Rats Scattering, Radiation Substrate Specificity Llovera, R.E. De Tullio, M. Alonso, L.G. Leissring, M.A. Kaufman, S.B. Roher, A.E. Gay, G.D.P. Morelli, L. Castaño, E.M. The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis |
| topic_facet |
Amines Binding energy Binding sites Colloids Diffusers (optical) Enzymes Flow interactions Forming Gelation Glycoproteins High performance liquid chromatography Hormones Insulin Mammals Mass spectrometry A stables After treatments Alzheimer diseases Catalytic domains Catalytic sites Complex formations Cortical tissues Degrading enzymes Diabetes Mellitus Genetic datums Limited proteolyses Mammalian brains Molar excesses Partial denaturations Rat brains Soluble fractions Substrate bindings Terminal fragments Integrodifferential equations amyloid beta protein dodecyl sulfate sodium insulinase recombinant enzyme Alzheimer disease amino terminal sequence article binding site complex formation controlled study enzyme active site enzyme denaturation human human tissue nonhuman pathogenesis priority journal protein conformation protein degradation protein domain protein folding protein protein interaction rat animal biological model brain chemistry enzyme specificity kinetics mass spectrometry metabolism pathology protein binding radiation scattering Mammalia Rattus Alzheimer Disease Amyloid beta-Protein Animals Binding Sites Brain Catalytic Domain Humans Insulysin Kinetics Mass Spectrometry Models, Biological Protein Binding Rats Scattering, Radiation Substrate Specificity |
| description |
Insulin-degrading enzyme (IDE) is central to the turnover of insulin and degrades amyloid β (Aβ) in the mammalian brain. Biochemical and genetic data support the notion that IDE may play a role in late onset Alzheimer disease (AD), and recent studies suggest an association between AD and diabetes mellitus type 2. Here we show that a natively folded recombinant IDE was capable of forming a stable complex with Aβ that resisted dissociation after treatment with strong denaturants. This interaction was also observed with rat brain IDE and detected in an SDS-soluble fraction from AD cortical tissue. Aβ sequence 17-27, known to be crucial in amyloid assembly, was sufficient to form a stable complex with IDE. Monomeric as opposed to aggregated Aβ was competent to associate irreversibly with IDE following a very slow kinetics (t1/2 ∼ 45 min). Partial denaturation of IDE as well as preincubation with a 10-fold molar excess of insulin prevented complex formation, suggesting that the irreversible interaction of Aβ takes place with at least part of the substrate binding site of the protease. Limited proteolysis showed that Aβ remained bound to a ∼25-kDa N-terminal fragment of IDE in an SDS-resistant manner. Mass spectrometry after in gel digestion of the IDE·Aβ complex showed that peptides derived from the region that includes the catalytic site of IDE were recovered with Aβ. Taken together, these results are suggestive of an unprecedented mechanism of conformation-dependent substrate binding that may perturb Aβ clearance, insulin turnover, and promote AD pathogenesis. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc. |
| format |
JOUR |
| author |
Llovera, R.E. De Tullio, M. Alonso, L.G. Leissring, M.A. Kaufman, S.B. Roher, A.E. Gay, G.D.P. Morelli, L. Castaño, E.M. |
| author_facet |
Llovera, R.E. De Tullio, M. Alonso, L.G. Leissring, M.A. Kaufman, S.B. Roher, A.E. Gay, G.D.P. Morelli, L. Castaño, E.M. |
| author_sort |
Llovera, R.E. |
| title |
The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis |
| title_short |
The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis |
| title_full |
The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis |
| title_fullStr |
The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis |
| title_full_unstemmed |
The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: Implications for Alzheimer disease pathogenesis |
| title_sort |
catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid β peptide: implications for alzheimer disease pathogenesis |
| url |
http://hdl.handle.net/20.500.12110/paper_00219258_v283_n25_p17039_Llovera |
| work_keys_str_mv |
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