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Isolation and characterization of mammalian D-aspartyl endopeptidase

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Summary.

The accumulation of D-isomers of aspartic acid (D-Asp) in proteins during aging has been implicated in the pathogenesis of Alzheimer’s disease (AD), cataracts and arteriosclerosis. Here, we identified a specific lactacystin-sensitive endopeptidase that cleaves the D-Asp-containing protein and named it D-aspartyl endopeptidase (DAEP). DAEP has a multi-complex structure (MW: 600 kDa) and is localized in the inner mitochondrial membrane. However, DAEP activity was not detected in E. coli, S. cerevisiae, and C. elegans. A specific inhibitor for DAEP, i-DAEP: (benzoyl-L-Arg-L-His-[D-Asp]-CH2Cl; MW: 563.01), was newly synthesized and inhibited DAEP activity (IC50, 3 µM), a factor of ten greater than lactacystin on DAEP. On the other hand, i-DAEP did not inhibit either the 20S or 26S proteasome. And we identified succinate dehydrogenase and glutamate dehydrogenase 1 as components of DAEP by affinity label using biotinylated i-DAEP. In the long life span of mammals, DAEP may serve as a scavenger against accumulation of racemized proteins in aging. Insights into DAEP will provide the foundation for developing treatments of diseases, such as AD, in which accumulation of D-Asp-containing proteins are implicated.

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References

  • DW Aswad (1984) ArticleTitleStoichiometric methylation of porcine adrenocorticotropin by protein carboxyl methyltransferase requires deamidation of asparagine 25. Evidence for methylation at the alpha-carboxyl group of atypical L-isoaspartyl residues J Biol Chem 259 10714–10721 Occurrence Handle6088513 Occurrence Handle1:CAS:528:DyaL2cXlvVKrsrY%3D

    PubMed  CAS  Google Scholar 

  • PA Cloos C Fledelius (2000) ArticleTitleCollagen fragments in urine derived from bone resorption are highly racemized and isomerized: a biological clock of protein aging with clinical potential Biochem J 345 473–480 Occurrence Handle10642504 Occurrence Handle1:CAS:528:DC%2BD3cXisFOlsbw%3D Occurrence Handle10.1042/0264-6021:3450473

    Article  PubMed  CAS  Google Scholar 

  • GH Fisher NM Garcia IL Payan R Cadilla-Perezrios WA Sheremata EH Man (1986) ArticleTitle D-aspartic acid in purified myelin and myelin basic protein Biochem Biophys Res Commun 135 683–687 Occurrence Handle2421722 Occurrence Handle1:CAS:528:DyaL28Xhs1eqsLw%3D Occurrence Handle10.1016/0006-291X(86)90047-1

    Article  PubMed  CAS  Google Scholar 

  • GH Fisher A D’Aniello A Vetere L Padula GP Cusano EH Man (1991) ArticleTitleFree D-aspartate and D-alanine in normal and Alzheimer brain Brain Res Bull 26 983–985 Occurrence Handle1933416 Occurrence Handle1:CAS:528:DyaK3MXltlegs7w%3D Occurrence Handle10.1016/0361-9230(91)90266-M

    Article  PubMed  CAS  Google Scholar 

  • N Fujii Y Ishibashi K Satoh M Fujino K Harada (1994) ArticleTitleSimultaneous racemization and isomerization at specific aspartic acid residues in alpha B-crystallin from the aged human lens Biochim Biophys Acta 1204 157–163 Occurrence Handle8142454 Occurrence Handle1:CAS:528:DyaK2cXhvVGluro%3D

    PubMed  CAS  Google Scholar 

  • T Geiger S Clarke (1987) ArticleTitleDeamidation, isomerization, and racemization at asparaginyl and aspartyl residues in peptides. Succinimide-linked reactions that contribute to protein degradation J Biol Chem 262 785–794 Occurrence Handle3805008 Occurrence Handle1:CAS:528:DyaL2sXksFyitQ%3D%3D

    PubMed  CAS  Google Scholar 

  • S Ishiura Y Mabuchi Y Urakami-Manaka K Isobe K Tagawa K Maruyama H Sorimachi K Suzuki (1997) Proteases involved in the processing of the Alzheimer’s disease amyloid precursor protein VK Hopsu-Havu M Järvinen H Kirschke (Eds) Proteolysis in cell functions IOS Press Amsterdam, Berlin, Oxford, Tokyo, Washington, DC 507–512

    Google Scholar 

  • A Iwamatsu N Yoshida-Kubomura (1996) ArticleTitleSystematic peptide fragmentation of polyvinylidene difluoride(PVDF)-immobilized proteins prior to microsequencing J Biochem (Tokyo) 120 29–34 Occurrence Handle1:CAS:528:DyaK28XltFGltrs%3D

    CAS  Google Scholar 

  • T Iwatsubo TC Saido DM Mann VM Lee JQ Trojanowski (1996) ArticleTitleFull-length amyloid-beta (1-42(43)) and amino-terminally modified and truncated amyloid-beta 42(43) deposit in diffuse plaques Am J Pathol 149 1823–1830 Occurrence Handle8952519 Occurrence Handle1:CAS:528:DyaK2sXjtFOntQ%3D%3D

    PubMed  CAS  Google Scholar 

  • HO Kanayama T Tamura S Ugai S Kagawa N Tanahashi T Yoshimura K Tanaka A Ichihara (1992) ArticleTitleDemonstration that a human 26S proteolytic complex consists of a proteasome and multiple associated protein components and hydrolyzes ATP and ubiquitin-ligated proteins by closely linked mechanisms Eur J Biochem 206 567–578 Occurrence Handle1317798 Occurrence Handle1:CAS:528:DyaK38XksVehsLc%3D Occurrence Handle10.1111/j.1432-1033.1992.tb16961.x

    Article  PubMed  CAS  Google Scholar 

  • A Kenessey SH Yen WK Liu XR Yang DS Dunlop (1995) ArticleTitleDetection of D-aspartate in tau proteins associated with Alzheimer paired helical filaments Brain Res 675 183–189 Occurrence Handle7796127 Occurrence Handle1:CAS:528:DyaK2MXksVylu7s%3D Occurrence Handle10.1016/0006-8993(95)00061-T

    Article  PubMed  CAS  Google Scholar 

  • T Kinouchi S Ishiura Y Mabuchi Y Urakami-Manaka H Nishio Y Nishiuchi M Tsunemi K Takada M Watanabe M Ikeda H Matsui S Tomioka H Kawahara T Hamamoto K Suzuki Y Kagawa (2004) ArticleTitleMammalian D-aspartyl endopeptidase: a scavenger for noxious racemized proteins in aging Biochem Biophys Res Commun 314 730–736 Occurrence Handle14741696 Occurrence Handle1:CAS:528:DC%2BD2cXltF2jtw%3D%3D Occurrence Handle10.1016/j.bbrc.2003.12.147

    Article  PubMed  CAS  Google Scholar 

  • JM Lee L Petrucelli G Fisher S Ramdath J Castillo MM Di Fiore A D’Aniello (2002) ArticleTitleEvidence for D-aspartyl-beta-amyloid secretase activity in human brain J Neuropathol Exp Neurol 61 125–131 Occurrence Handle11855381 Occurrence Handle1:CAS:528:DC%2BD38XitFOku70%3D

    PubMed  CAS  Google Scholar 

  • EH Man ME Sandhouse J Burg GH Fisher (1983) ArticleTitleAccumulation of D-aspartic acid with age in the human brain Science 220 1407–1408 Occurrence Handle6857259 Occurrence Handle1:CAS:528:DyaL3sXktlKnu7c%3D Occurrence Handle10.1126/science.6857259

    Article  PubMed  CAS  Google Scholar 

  • PN McFadden S Clarke (1982) ArticleTitleMethylation at D-aspartyl residues in erythrocytes: possible step in the repair of aged membrane proteins Proc Natl Acad Sci USA 79 2460–2464 Occurrence Handle6123997 Occurrence Handle1:CAS:528:DyaL38Xhslylu7k%3D Occurrence Handle10.1073/pnas.79.8.2460

    Article  PubMed  CAS  Google Scholar 

  • H Mori K Ishii T Tomiyama Y Furiya N Sahara S Asano N Endo T Shirasawa K Takio (1994) ArticleTitleRacemization: its biological significance on neuropathogenesis of Alzheimer’s disease Tohoku J Exp Med 174 251–262 Occurrence Handle7761990 Occurrence Handle1:CAS:528:DyaK2MXks1Sitr4%3D Occurrence Handle10.1620/tjem.174.251

    Article  PubMed  CAS  Google Scholar 

  • S Muraoka N Fujii I Tamanoi K Harada (1987) ArticleTitleCharacterization of a protein containing D-aspartic acid in aged mouse lens Biochem Biophys Res Commun 146 1432–1438 Occurrence Handle3619938 Occurrence Handle1:CAS:528:DyaL2sXmtlWltLY%3D Occurrence Handle10.1016/0006-291X(87)90810-2

    Article  PubMed  CAS  Google Scholar 

  • Y Nagata A Fukuda M Sakai T Iida K Kawaguchi-Nagata (2001) ArticleTitle D-Amino acid contents of mitochondria and some purple bacteria J Mol Catal B 12 109–113 Occurrence Handle1:CAS:528:DC%2BD3MXht1Oqurs%3D Occurrence Handle10.1016/S1381-1177(00)00210-1

    Article  CAS  Google Scholar 

  • S Omura H Ikeda H Tanaka (1991) ArticleTitleSelective production of specific components of avermectins in Streptomyces avermitilis J Antibiot (Tokyo) 44 560–563 Occurrence Handle1:STN:280:By6B1cngs1Q%3D

    CAS  Google Scholar 

  • JT Powell N Vine M Crossman (1992) ArticleTitleOn the accumulation of D-aspartate in elastin and other proteins of the ageing aorta Atherosclerosis 97 201–208 Occurrence Handle1466664 Occurrence Handle1:CAS:528:DyaK3sXht1Wmt7k%3D Occurrence Handle10.1016/0021-9150(92)90132-Z

    Article  PubMed  CAS  Google Scholar 

  • M Seeger C Gordon K Ferrell W Dubiel (1996) ArticleTitleCharacteristics of 26S proteases from fission yeast mutants, which arrest in mitosis J Mol Biol 263 423–431 Occurrence Handle8918598 Occurrence Handle1:CAS:528:DyaK28Xms1yrs70%3D Occurrence Handle10.1006/jmbi.1996.0586

    Article  PubMed  CAS  Google Scholar 

  • R Shapira GE Austin SS Mirra (1988) ArticleTitleNeuritic plaque amyloid in Alzheimer’s disease is highly racemized J Neurochem 50 69–74 Occurrence Handle3121789 Occurrence Handle1:CAS:528:DyaL1cXltlOkug%3D%3D Occurrence Handle10.1111/j.1471-4159.1988.tb02960.x

    Article  PubMed  CAS  Google Scholar 

  • SD Shapiro SK Endicott MA Province JA Pierce EJ Campbell (1991) ArticleTitleMarked longevity of human lung parenchymal elastic fibers deduced from prevalence of D-aspartate and nuclear weapons-related radiocarbon J Clin Invest 87 1828–1834 Occurrence Handle2022748 Occurrence Handle1:CAS:528:DyaK3MXisVCqtLs%3D Occurrence Handle10.1172/JCI115204

    Article  PubMed  CAS  Google Scholar 

  • T Shimizu A Watanabe M Ogawara H Mori T Shirasawa (2000) ArticleTitleIsoaspartate formation and neurodegeneration in Alzheimer’s disease Arch Biochem Biophys 381 225–234 Occurrence Handle11032409 Occurrence Handle1:CAS:528:DC%2BD3cXmsFems78%3D Occurrence Handle10.1006/abbi.2000.1955

    Article  PubMed  CAS  Google Scholar 

  • T Tomiyama S Asano Y Furiya T Shirasawa N Endo H Mori (1994) ArticleTitleRacemization of Asp23 residue affects the aggregation properties of Alzheimer amyloid beta protein analogues J Biol Chem 269 10205–10208 Occurrence Handle8144598 Occurrence Handle1:CAS:528:DyaK2cXjt1Slsbk%3D

    PubMed  CAS  Google Scholar 

  • T Watanabe Y Kera T Matsumoto RH Yamada (1996) ArticleTitlePurification and kinetic properties of a D-amino-acid peptide hydrolyzing enzyme from pig kidney cortex and its tentative identification with renal membrane dipeptidase Biochim Biophys Acta 1298 109–118 Occurrence Handle8948495

    PubMed  Google Scholar 

  • DJ Weber PN McFadden B Caughey (1998) ArticleTitleMeasurement of altered aspartyl residues in the scrapie associated form of prion protein Biochem Biophys Res Commun 246 606–608 Occurrence Handle9618258 Occurrence Handle1:CAS:528:DyaK1cXjsFyjtLs%3D Occurrence Handle10.1006/bbrc.1998.8672

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Radiation Life Science and Radiation Medical Science, Research Reactor Institute, Kyoto University, Osaka, Japan

    T. Kinouchi & N. Fujii

  2. Peptide Institute, Inc., Osaka, Japan

    H. Nishio, Y. Nishiuchi, M. Tsunemi & K. Takada

  3. Department of Biochemistry, Jichi Medical School, Tochigi, Japan

    T. Hamamoto

  4. Department of Medical Chemistry, Women’s University of Nutrition, Saitama, Japan

    Y. Kagawa

Authors
  1. T. Kinouchi
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  2. H. Nishio
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  3. Y. Nishiuchi
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  4. M. Tsunemi
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  5. K. Takada
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  6. T. Hamamoto
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  7. Y. Kagawa
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  8. N. Fujii
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Kinouchi, T., Nishio, H., Nishiuchi, Y. et al. Isolation and characterization of mammalian D-aspartyl endopeptidase. Amino Acids 32, 79–85 (2007). https://doi.org/10.1007/s00726-006-0348-4

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  • DOI: https://doi.org/10.1007/s00726-006-0348-4

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