Processing of APP Directory UMM :Data Elmu:jurnal:B:Brain Research:Vol886.Issue1-2.Nov2000:

R .L. Neve et al. Brain Research 886 2000 54 –66 55 [118]. Cells expressing FAD mutants of APP and the transmission [28]. There has been some question of presenilins are reported to secrete increased amounts of whether C100 exerts its neurotoxic effects from the inside Ab , suggesting a link of this variant of Ab to AD or the outside of the cell [23,117]. Our data of the past 6 1 – 42 pathogenesis. Consequently, a leading hypothesis for the years suggest strongly that C100 kills from inside the cell; etiology of AD is that increased Ab is a shared this is supported by the observation that C100 is not 1 – 42 molecular correlate of FAD mutations, and that it repre- secreted, even when it carries a signal peptide [19,14,66]. sents a gain of deleterious function that can cause FAD Although at least one group has reported neurotoxicity due [38] and may be an essential early event in AD [118]. to the addition of C100 to the culture medium [50], we While this ‘amyloid hypothesis’ is attractive, molecular believe that that type of neurotoxicity is mechanistically mechanisms other than those mediated by extracellular Ab different from the neurodegeneration that we observe upon could also lead to AD neurodegeneration. expression of C100 within primary neurons. These mechanisms are likely to be linked in some way The findings that APP interacts with the signaling to the b-amyloid protein precursor APP, the source of molecule G , that FAD mutants of APP can cause G - o o Ab. One of the most compelling pieces of evidence that mediated apoptosis in neuronal cells, and that these same links AD neurodegeneration to APP and or its Ab-con- FAD mutants of APP cause the intracellular accumulation taining derivatives is the early finding that the APP gene is of C100, suggested to us the following working hypoth- on chromosome 21: virtually all individuals trisomic for esis: In the brain a portion of APP is present as an integral this chromosome show AD-like neuropathology by the age plasma membrane protein that mediates the transduction of of 40. Additionally, it has been discovered that specific extracellular signals into the cell via its C-terminal tail, and mutations in APP cause some forms of familial FAD. abnormal accumulation of its Ab-containing C-terminal These data have raised the possibility that AD may result tail in the neuron causes progressive dysfunction of APP from an alteration in the normal function of APP [74,76], signaling in AD, resulting in apoptosis. This hypothesis has and have refocused attention on the delineation of the been supported by the finding that the intracellular C- function that APP subserves in the brain. It has been terminal tail of APP interacts with the cell cycle protein shown [47,83] that in the brain a percentage of APP is APP-BP1 [10,9], and with members of the Fe65 family of present on the cell surface, and it is proposed [76,83] that adaptor proteins reviewed in Ref. [89]. Additional sup- this cell surface APP mediates the transduction of extracel- port for this hypothesis emerged with the recent report [60] lular signals into the cell via its C-terminal tail. that the C31 peptide of APP, which is derived from C100 Nishimoto and his colleagues [75] showed that APP and within which are contained the binding sites for the binds to the brain-specific signal transducing G protein G ; above proteins, is elevated in AD brain and is a potent o independent confirmation of this finding has subsequently inducer of apoptosis. been published [4,5]. It was then discovered [113] that V642 ‘London’ FAD mutants of APP induce neuronal DNA fragmentation, a feature of apoptosis, in a neuronal

2. Processing of APP

cell line. This fragmentation is independent of Ab 1 – 42 production [114] and is mediated by the Gb g complex of Most of what we know about APP processing has come 2 2 G [29]. These data support the notion that APP has an from work with cultured cells. APP matures through the o intrinsic signaling function in the neuron, which becomes constitutive secretory pathway. Some fraction of the APP ligand-independent when APP is mutated at V642. is endoproteolytically cleaved at the cell surface within the To examine the mechanism by which FAD APP might Ab sequence by the a-secretase, which generates the cause apoptosis in neurons, we [66] expressed five differ- neuroprotective secreted amyloid precursor protein ent Alzheimer mutations of APP in primary neurons via APP and nonamyloidogenic 3 kDa Ab secreted prod- sa recombinant herpes simplex virus HSV vectors, and ucts [81,37,65]. APP is readily detected in human plasma Sa quantified the levels of APP metabolites. The predominant and cerebrospinal fluid. intracellular accumulation product was a C-terminal frag- Endocytosis of cell surface APP generates C-terminal ment of APP that co-migrated with the protein product of fragments of 8–12 kDa that are degraded in the lysosomes an HSV recombinant expressing the C-terminal 100 amino [31,36]; some of these, generated by b-secretase cleavage acids C100 of APP. Interestingly, we had proposed at the N-terminus of the Ab sequence analogous to C100, previously that C100 is involved in the etiology of are amyloidogenic. A second protease activity called g- Alzheimer disease [73]. It is neurotoxic in vitro secretase cleaves these C-terminal fragments of APP to [116,24,97,101,57] and is amyloidogenic [17– release the full-length Ab protein. The b-secretase has 19,64,111,27,105]. In addition, expression of C100 in vivo been identified [107,42,96,115], and accumulating data can cause neuropathology that is similar in some ways to suggest strongly that presenilin is the g-secretase [58]. that in AD, including neurodegeneration and cognitive Thus, both Ab and C100-like fragments of APP are dysfunction [3,25,48,71,72,80,91,104], as well as increases produced normally at low levels by the cell. Most of the in acetylcholinesterase [92] and abnormalities in synaptic Ab that is generated is secreted, although small amounts 56 R can be detected intracellularly. C100-like amyloidogenic and colleagues have shown that b-secretase cleavage fragments have been detected only intracellularly. products of APP are present in fetal and neonatal Down It is important to note that APP processing is cell syndrome brain at twice normal levels [93] and personal type-specific. LeBlanc and colleagues have reported that communication with Dr. Russo. We have hypothesized human neurons secrete more 4 kDa than 3 kDa Ab, and that abnormal accumulation of the Ab-containing C-termi- metabolize approximately 40 of newly-synthesized APP nal tail C100 of APP in neurons also occurs in Alzheimer through the a-secretase pathway [53,55]. Moreover, disease. human neurons produce five C-terminal fragments of APP in a pattern seen uniquely in human brain [21,53]. The two largest C-terminal derivatives have the entire Ab sequence

3. APP as a signaling molecule