Role of ERAB/L-3-hydroxyacyl-coenzyme A dehydrogenase type II activity in Aβ-induced cytotoxicity

Shi Du Yan, Yigong Shi, Aiping Zhu, Jin Fu, Huaijie Zhu, Yucui Zhu, Lenneen Gibson, Eric Stern, Kate Collison, Futwan Al-Mohanna, Satoshi Ogawa, Alex Roher, Steven G. Clarke, David Stern

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Abstract

Endoplasmic reticulum-associated amyloid β-peptide (Aβ)-binding protein (ERAB)/L-3-hydroxyacyl-CoA dehydrogenase type II (HADH II) is expressed at high levels in Alzheimer's disease (AD)-affected brain, binds Aβ, and contributes to Aβ-induced cytotoxicity. Purified recombinant ERAB/HADH II catalyzed the NADH-dependent reduction of S-acetoacetyl-CoA with a K(m) of ≃68 μM and a V(max) of ≃430 μmol/min/mg. The contribution of ERAB/HADH II enzymatic activity to Aβ-mediated cellular dysfunction was studied by site-directed mutagenesis in the catalytic domain (Y168G/K172G). Although COS cells cotransfected to overexpress wild-type ERAB/HADH II and variant β-amyloid precursor protein (βAPP(V717G)) showed DNA fragmentation, cotransfection with Y168G/K172G-altered ERAB and βAPP(V717G) was without effect. We thus asked whether the enzyme might recognize alcohol substrates of which the aldehyde products could be cytotoxic; ERAB/HADH II catalyzed oxidation of a variety of simple alcohols (C2-C10) to their respective aldehydes in the presence of NAD+ and NAD-dependent oxidation of 17β- estradiol. Addition of micromolar levels of synthetic Aβ(1-40) to purified ERAB/HADH II inhibited, in parallel, reduction of S-acetoacetyl-Co(A) (K(i) ≃ 1.6 μM), as well as oxidation of 17β-estradiol (K(i) ≃ 3.2 μM) and (- )-2-octanol (K(i) ≃ 2.6 μM). Because micromolar levels of Aβ were required to inhibit ERAB/HADH II activity, whereas Aβ binding to ERAB/HADH II occurred at much lower concentrations (K(m) ≃ 40-70) nM) 40-70 the latter more closely simulating Aβ levels within cells, Aβ perturbation of ERAB/HADH II was likely to result from mechanisms other than the direct modulation of enzymatic activity. Cells cotransfected to overexpress ERAB/HADH II and βAPP(V717G) generated malondialdehyde-protein and 4- hydroxynonenal-protein epitopes, which were detectable only at the lowest levels in cells overexpressing either ERAB/HADH II or βAPP(V717G) alone. Generation of such toxic aldehydes was not observed in cells contransfected to overexpress Y168G/K172G-altered ERAB and βAPP(V717G). We conclude that the generalized alcohol dehydrogenase activity of ERAB/HADH II is central to the cytotoxicity observed in an Aβ-rich environment.

Original languageEnglish (US)
Pages (from-to)2145-2156
Number of pages12
JournalJournal of Biological Chemistry
Volume274
Issue number4
DOIs
StatePublished - Jan 22 1999

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Coenzyme A
Cytotoxicity
Oxidoreductases
Aldehydes
NAD
Oxidation
Estradiol
3-Hydroxyacyl-CoA Dehydrogenase
Alcohols
Mutagenesis
Amyloid beta-Protein Precursor
Alcohol Dehydrogenase
Poisons
COS Cells
DNA Fragmentation
Site-Directed Mutagenesis
Malondialdehyde
Amyloid
Endoplasmic Reticulum
Epitopes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Role of ERAB/L-3-hydroxyacyl-coenzyme A dehydrogenase type II activity in Aβ-induced cytotoxicity. / Yan, Shi Du; Shi, Yigong; Zhu, Aiping; Fu, Jin; Zhu, Huaijie; Zhu, Yucui; Gibson, Lenneen; Stern, Eric; Collison, Kate; Al-Mohanna, Futwan; Ogawa, Satoshi; Roher, Alex; Clarke, Steven G.; Stern, David.

In: Journal of Biological Chemistry, Vol. 274, No. 4, 22.01.1999, p. 2145-2156.

Research output: Contribution to journalArticle

Yan, SD, Shi, Y, Zhu, A, Fu, J, Zhu, H, Zhu, Y, Gibson, L, Stern, E, Collison, K, Al-Mohanna, F, Ogawa, S, Roher, A, Clarke, SG & Stern, D 1999, 'Role of ERAB/L-3-hydroxyacyl-coenzyme A dehydrogenase type II activity in Aβ-induced cytotoxicity', Journal of Biological Chemistry, vol. 274, no. 4, pp. 2145-2156. https://doi.org/10.1074/jbc.274.4.2145
Yan, Shi Du ; Shi, Yigong ; Zhu, Aiping ; Fu, Jin ; Zhu, Huaijie ; Zhu, Yucui ; Gibson, Lenneen ; Stern, Eric ; Collison, Kate ; Al-Mohanna, Futwan ; Ogawa, Satoshi ; Roher, Alex ; Clarke, Steven G. ; Stern, David. / Role of ERAB/L-3-hydroxyacyl-coenzyme A dehydrogenase type II activity in Aβ-induced cytotoxicity. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 4. pp. 2145-2156.
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abstract = "Endoplasmic reticulum-associated amyloid β-peptide (Aβ)-binding protein (ERAB)/L-3-hydroxyacyl-CoA dehydrogenase type II (HADH II) is expressed at high levels in Alzheimer's disease (AD)-affected brain, binds Aβ, and contributes to Aβ-induced cytotoxicity. Purified recombinant ERAB/HADH II catalyzed the NADH-dependent reduction of S-acetoacetyl-CoA with a K(m) of ≃68 μM and a V(max) of ≃430 μmol/min/mg. The contribution of ERAB/HADH II enzymatic activity to Aβ-mediated cellular dysfunction was studied by site-directed mutagenesis in the catalytic domain (Y168G/K172G). Although COS cells cotransfected to overexpress wild-type ERAB/HADH II and variant β-amyloid precursor protein (βAPP(V717G)) showed DNA fragmentation, cotransfection with Y168G/K172G-altered ERAB and βAPP(V717G) was without effect. We thus asked whether the enzyme might recognize alcohol substrates of which the aldehyde products could be cytotoxic; ERAB/HADH II catalyzed oxidation of a variety of simple alcohols (C2-C10) to their respective aldehydes in the presence of NAD+ and NAD-dependent oxidation of 17β- estradiol. Addition of micromolar levels of synthetic Aβ(1-40) to purified ERAB/HADH II inhibited, in parallel, reduction of S-acetoacetyl-Co(A) (K(i) ≃ 1.6 μM), as well as oxidation of 17β-estradiol (K(i) ≃ 3.2 μM) and (- )-2-octanol (K(i) ≃ 2.6 μM). Because micromolar levels of Aβ were required to inhibit ERAB/HADH II activity, whereas Aβ binding to ERAB/HADH II occurred at much lower concentrations (K(m) ≃ 40-70) nM) 40-70 the latter more closely simulating Aβ levels within cells, Aβ perturbation of ERAB/HADH II was likely to result from mechanisms other than the direct modulation of enzymatic activity. Cells cotransfected to overexpress ERAB/HADH II and βAPP(V717G) generated malondialdehyde-protein and 4- hydroxynonenal-protein epitopes, which were detectable only at the lowest levels in cells overexpressing either ERAB/HADH II or βAPP(V717G) alone. Generation of such toxic aldehydes was not observed in cells contransfected to overexpress Y168G/K172G-altered ERAB and βAPP(V717G). We conclude that the generalized alcohol dehydrogenase activity of ERAB/HADH II is central to the cytotoxicity observed in an Aβ-rich environment.",
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T1 - Role of ERAB/L-3-hydroxyacyl-coenzyme A dehydrogenase type II activity in Aβ-induced cytotoxicity

AU - Yan, Shi Du

AU - Shi, Yigong

AU - Zhu, Aiping

AU - Fu, Jin

AU - Zhu, Huaijie

AU - Zhu, Yucui

AU - Gibson, Lenneen

AU - Stern, Eric

AU - Collison, Kate

AU - Al-Mohanna, Futwan

AU - Ogawa, Satoshi

AU - Roher, Alex

AU - Clarke, Steven G.

AU - Stern, David

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N2 - Endoplasmic reticulum-associated amyloid β-peptide (Aβ)-binding protein (ERAB)/L-3-hydroxyacyl-CoA dehydrogenase type II (HADH II) is expressed at high levels in Alzheimer's disease (AD)-affected brain, binds Aβ, and contributes to Aβ-induced cytotoxicity. Purified recombinant ERAB/HADH II catalyzed the NADH-dependent reduction of S-acetoacetyl-CoA with a K(m) of ≃68 μM and a V(max) of ≃430 μmol/min/mg. The contribution of ERAB/HADH II enzymatic activity to Aβ-mediated cellular dysfunction was studied by site-directed mutagenesis in the catalytic domain (Y168G/K172G). Although COS cells cotransfected to overexpress wild-type ERAB/HADH II and variant β-amyloid precursor protein (βAPP(V717G)) showed DNA fragmentation, cotransfection with Y168G/K172G-altered ERAB and βAPP(V717G) was without effect. We thus asked whether the enzyme might recognize alcohol substrates of which the aldehyde products could be cytotoxic; ERAB/HADH II catalyzed oxidation of a variety of simple alcohols (C2-C10) to their respective aldehydes in the presence of NAD+ and NAD-dependent oxidation of 17β- estradiol. Addition of micromolar levels of synthetic Aβ(1-40) to purified ERAB/HADH II inhibited, in parallel, reduction of S-acetoacetyl-Co(A) (K(i) ≃ 1.6 μM), as well as oxidation of 17β-estradiol (K(i) ≃ 3.2 μM) and (- )-2-octanol (K(i) ≃ 2.6 μM). Because micromolar levels of Aβ were required to inhibit ERAB/HADH II activity, whereas Aβ binding to ERAB/HADH II occurred at much lower concentrations (K(m) ≃ 40-70) nM) 40-70 the latter more closely simulating Aβ levels within cells, Aβ perturbation of ERAB/HADH II was likely to result from mechanisms other than the direct modulation of enzymatic activity. Cells cotransfected to overexpress ERAB/HADH II and βAPP(V717G) generated malondialdehyde-protein and 4- hydroxynonenal-protein epitopes, which were detectable only at the lowest levels in cells overexpressing either ERAB/HADH II or βAPP(V717G) alone. Generation of such toxic aldehydes was not observed in cells contransfected to overexpress Y168G/K172G-altered ERAB and βAPP(V717G). We conclude that the generalized alcohol dehydrogenase activity of ERAB/HADH II is central to the cytotoxicity observed in an Aβ-rich environment.

AB - Endoplasmic reticulum-associated amyloid β-peptide (Aβ)-binding protein (ERAB)/L-3-hydroxyacyl-CoA dehydrogenase type II (HADH II) is expressed at high levels in Alzheimer's disease (AD)-affected brain, binds Aβ, and contributes to Aβ-induced cytotoxicity. Purified recombinant ERAB/HADH II catalyzed the NADH-dependent reduction of S-acetoacetyl-CoA with a K(m) of ≃68 μM and a V(max) of ≃430 μmol/min/mg. The contribution of ERAB/HADH II enzymatic activity to Aβ-mediated cellular dysfunction was studied by site-directed mutagenesis in the catalytic domain (Y168G/K172G). Although COS cells cotransfected to overexpress wild-type ERAB/HADH II and variant β-amyloid precursor protein (βAPP(V717G)) showed DNA fragmentation, cotransfection with Y168G/K172G-altered ERAB and βAPP(V717G) was without effect. We thus asked whether the enzyme might recognize alcohol substrates of which the aldehyde products could be cytotoxic; ERAB/HADH II catalyzed oxidation of a variety of simple alcohols (C2-C10) to their respective aldehydes in the presence of NAD+ and NAD-dependent oxidation of 17β- estradiol. Addition of micromolar levels of synthetic Aβ(1-40) to purified ERAB/HADH II inhibited, in parallel, reduction of S-acetoacetyl-Co(A) (K(i) ≃ 1.6 μM), as well as oxidation of 17β-estradiol (K(i) ≃ 3.2 μM) and (- )-2-octanol (K(i) ≃ 2.6 μM). Because micromolar levels of Aβ were required to inhibit ERAB/HADH II activity, whereas Aβ binding to ERAB/HADH II occurred at much lower concentrations (K(m) ≃ 40-70) nM) 40-70 the latter more closely simulating Aβ levels within cells, Aβ perturbation of ERAB/HADH II was likely to result from mechanisms other than the direct modulation of enzymatic activity. Cells cotransfected to overexpress ERAB/HADH II and βAPP(V717G) generated malondialdehyde-protein and 4- hydroxynonenal-protein epitopes, which were detectable only at the lowest levels in cells overexpressing either ERAB/HADH II or βAPP(V717G) alone. Generation of such toxic aldehydes was not observed in cells contransfected to overexpress Y168G/K172G-altered ERAB and βAPP(V717G). We conclude that the generalized alcohol dehydrogenase activity of ERAB/HADH II is central to the cytotoxicity observed in an Aβ-rich environment.

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