Article

Degradation of the amyloid beta-protein by the novel mitochondrial peptidasome, PreP

Department of Biochemistry and Biophysics , Stockholm University, Tukholma, Stockholm, Sweden
Journal of Biological Chemistry (Impact Factor: 4.6). 10/2006; 281(39):29096-104. DOI: 10.1074/jbc.M602532200
Source: PubMed

ABSTRACT Recently we have identified the novel mitochondrial peptidase responsible for degrading presequences and other short unstructured peptides in mitochondria, the presequence peptidase, which we named PreP peptidasome. In the present study we have identified and characterized the human PreP homologue, hPreP, in brain mitochondria, and we show its capacity to degrade the amyloid beta-protein (Abeta). PreP belongs to the pitrilysin oligopeptidase family M16C containing an inverted zinc-binding motif. We show that hPreP is localized to the mitochondrial matrix. In situ immuno-inactivation studies in human brain mitochondria using anti-hPreP antibodies showed complete inhibition of proteolytic activity against Abeta. We have cloned, overexpressed, and purified recombinant hPreP and its mutant with catalytic base Glu(78) in the inverted zinc-binding motif replaced by Gln. In vitro studies using recombinant hPreP and liquid chromatography nanospray tandem mass spectrometry revealed novel cleavage specificities against Abeta-(1-42), Abeta-(1-40), and Abeta Arctic, a protein that causes increased protofibril formation an early onset familial variant of Alzheimer disease. In contrast to insulin degrading enzyme, which is a functional analogue of hPreP, hPreP does not degrade insulin but does degrade insulin B-chain. Molecular modeling of hPreP based on the crystal structure at 2.1 A resolution of AtPreP allowed us to identify Cys(90) and Cys(527) that form disulfide bridges under oxidized conditions and might be involved in redox regulation of the enzyme. Degradation of the mitochondrial Abeta by hPreP may potentially be of importance in the pathology of Alzheimer disease.

0 Followers
 · 
101 Views
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Mitochondrial dysfunction is an important factor in the pathogenesis of age-related diseases, including neurodegenerative diseases like Alzheimer's and Parkinson's spectrum disorders. A polymorphism in Translocase of the Outer Mitochondrial Membrane - 40 kD (TOMM40) is associated with risk and age-of onset of late-onset AD, and is the only nuclear- encoded gene identified in genetic studies to date that presumably contributes to LOAD-related mitochondria dysfunction. In this review, we describe the TOM40-mediated mitochondrial protein import mechanism, and discuss the evidence linking TOM40 with Alzheimer's (AD) and Parkinson's (PD) diseases. All but 36 of the >~1,500 mitochondrial proteins are encoded by the nucleus and are synthesized on cytoplasmic ribosomes, and most of these are imported into mitochondria through the TOM complex, of which TOM40 is the central pore, mediating communication between the cytoplasm and the mitochondrial interior. APP enters and obstructs the TOM40 pore, inhibiting import of OXPHOS-related proteins and disrupting the mitochondrial redox balance. Other pathogenic proteins, such as Aβ and alpha-synuclein, readily pass through the pore and cause toxic effects by directly inhibiting mitochondrial enzymes. Healthy mitochondria normally import and degrade the PD-related protein Pink1, but Pink1 exits mitochondria if the membrane potential collapses and initiates Parkin-mediated mitophagy. Under normal circumstances, this process helps clear dysfunctional mitochondria and contributes to cellular health, but PINK1 mutations associated with PD exit mitochondria with intact membrane potentials, disrupting mitochondrial dynamics, leading to pathology. Thus, TOM40 plays a central role in the mitochondrial dysfunction that underlies age-related neurodegenerative diseases. Learning about the factors that control TOM40 levels and activity, and how TOM40, specifically, and the TOM complex, generally, interacts with potentially pathogenic proteins, will provide deeper insights to AD and PD pathogenesis, and possibly new targets for preventative and/or therapeutic treatments.
  • [Show abstract] [Hide abstract]
    ABSTRACT: Significance: Mitochondria are complex dynamic organelles pivotal for cellular physiology and human health. Failure to maintain mitochondrial health leads to numerous maladies that include late-onset neurodegenerative diseases and cardiovascular disorders. Furthermore, a decline in mitochondrial health is prevalent with aging. A set of evolutionary conserved mechanisms known as mitochondrial quality control (MQC) is involved in recognition and correction of the mitochondrial proteome. Recent Advances: Here, we review current knowledge and latest developments in MQC. We particularly focus on the proteolytic aspect of MQC and its impact on health and aging. Critical Issues: While our knowledge about MQC is steadily growing, critical gaps remain in the mechanistic understanding of how MQC modules sense damage and preserve mitochondrial welfare, particularly in higher organisms. Future Directions: Delineating how coordinated action of the MQC modules orchestrates physiological responses on both organellar and cellular levels will further elucidate the current picture of MQC's role and function in health, cellular stress and degenerative diseases.
    Antioxidants and Redox Signaling 12/2014; DOI:10.1089/ars.2014.6199 · 7.67 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Alzheimer ' s disease (AD) represents the most common form of dementia among people age 65 and older, affecting more than 35 million people worldwide and representing 50–56% of cases at autopsy and in clinical series. Clinically, AD is characterized by a progressive cognitive deteriora-tion, together with impairments in behavior, language, and visuospatial skills, culminating in the premature death of the individual typically 3–9 years after diagnosis (Querfurth and LaFerla 2010, 329). The great majority of AD cases are sporadic in origin with a late onset, while a small propor-tion (< 1%) has genetic origin and involves mutations in amyloid β protein precursor (AβPP) and presenilins 1 and 2 (PS1 and PS2), leading to auto-somal dominant familial AD with an early onset. Additionally, the allelic abnormalities of the apolipoprotein E (APOE) gene on chromosome 19 are responsible for both anticipated onset and increase in severity of inherited and sporadic AD (Rocchi et al. 2003, 1). Neuropathologically, AD has as main hallmarks the selective neuronal and synaptic loss, the deposition of.indd 89 12/17/10 1:24:22 PM 12/17/10 1:24:22 PM
    Dementia Volume II: Science and Biology, Edited by Patrick McNamara, 01/2010: chapter 3: pages 89-114; ABC-CLIO.