Determination of cleavage site of Reelin between its sixth and seventh repeat and contribution of meprin metalloproteases to the cleavage

Sato, Yoshitaka; Kobayashi, Daichi; Kohno, Takao; Kidani, Yujiro; Prox, Johannes; Becker‐Pauly, Christoph; Hattori, Mitsuharu

doi:10.1093/jb/mvv102

Cited by 26 publications

(31 citation statements)

References 44 publications

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“…WC cleavage occurs between Arg3455 and Ser3456, which is only six amino acid residues from the C-terminus and does not affect the apparent mobility in SDS-PAGE 14 . N-t cleavage dramatically decreases Dab1 phosphorylation by Reelin 30,31 , while C-t or WC cleavage modulate it 14,32 .…”

Section: Introductionmentioning

confidence: 95%

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Physiological significance of proteolytic processing of Reelin revealed by cleavage-resistant Reelin knock-in mice

Okugawa

Ogino

Shigenobu

et al. 2020

Preprint

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Reelin is a secreted protein that plays versatile roles in neuronal development and function, and hypoactivity of Reelin is implicated in many neuropsychiatric disorders. The strength of Reelin signaling is regulated by proteolytic processing, but its importance in vivo is not yet fully understood. Here, we generated Reelin knock-in (PA-DV KI) mice in which the key cleavage site of Reelin was abolished by mutation. As expected, the cleavage of Reelin was severely abrogated in the cerebral cortex and hippocampus of PA-DV KI mice. The amount of Dab1, whose degradation is induced by Reelin signaling, decreased in these tissues, indicating that the signaling strength of Reelin was augmented. The brains of PA-DV KI mice were largely structurally normal, but unexpectedly, the hippocampal layer was disturbed. This phenotype was ameliorated in hemizygote PA-DV KI mice, indicating that excess Reelin signaling is detrimental to hippocampal layer formation. The neuronal dendrites of PA-DV KI mice had more branches and were elongated compared to wild-type mice. These results present the first direct evidence of the physiological importance of Reelin cleavage and suggest that inhibition of Reelin cleavage would counteract neuropsychiatric disorders without causing severe systemic side effects.

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Section: Introductionmentioning

confidence: 95%

“…C-t cleavage occurs between Ala2688 and Asp2689, resulting in two fragments with molecular masses of 330 kDa (NR6) and 100 kDa ( Fig. 1A) 32 . WC cleavage occurs between Arg3455 and Ser3456, which is only six amino acid residues from the C-terminus and does not affect the apparent mobility in SDS-PAGE 14 .…”

Section: Introductionmentioning

confidence: 99%

Physiological significance of proteolytic processing of Reelin revealed by cleavage-resistant Reelin knock-in mice

Okugawa

Ogino

Shigenobu

et al. 2020

Preprint

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“…C-terminus and does not affect the apparent mobility in SDS-PAGE 14 . N-t cleavage dramatically decreases Dab1 phosphorylation by Reelin 30,31 , while C-t or WC cleavage modulate it 14,32 .…”

mentioning

confidence: 95%

“…C-t cleavage occurs between Ala2688 and Asp2689, resulting in two fragments with molecular masses of 330 kDa (NR6) and 100 kDa (Fig. 1A) 32 . WC cleavage occurs between Arg3455 and Ser3456, which is only six amino acid residues from the open Scientific RepoRtS | (2020) 10:4471 | https://doi.org/10.1038/s41598-020-61380-w www.nature.com/scientificreports www.nature.com/scientificreports/ Figure 1.…”

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confidence: 99%

Physiological significance of proteolytic processing of Reelin revealed by cleavage-resistant Reelin knock-in mice

Okugawa

Ogino

Shigenobu

et al. 2020

Sci Rep

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Reelin is a secreted protein that plays versatile roles in neuronal development and function. The strength of Reelin signaling is regulated by proteolytic processing, but its importance in vivo is not yet fully understood. Here, we generated Reelin knock-in (PA-DV KI) mice in which the key cleavage site of Reelin was abolished by mutation. As expected, the cleavage of Reelin was severely abrogated in the cerebral cortex and hippocampus of PA-DV KI mice. The amount of Dab1, whose degradation is induced by Reelin signaling, decreased in these tissues, indicating that the signaling strength of Reelin was augmented. The brains of PA-DV KI mice were largely structurally normal, but unexpectedly, the hippocampal layer was disturbed. This phenotype was ameliorated in hemizygote PA-DV KI mice, indicating that excess Reelin signaling is detrimental to hippocampal layer formation. The neuronal dendrites of PA-DV KI mice had more branches and were elongated compared to wild-type mice. These results present the first direct evidence of the physiological importance of Reelin cleavage.Reelin is a large secreted glycoprotein that regulates many important events in mammalian brain development 1-3 . It is also involved in synaptic plasticity and the modulation of higher brain functions 1,4-8 . At the cellular level, Reelin induces the clustering of apolipoprotein E receptor 2 (ApoER2) and very-low-density lipoprotein receptor (VLDLR), which is the prerequisite for the phosphorylation of the cytosolic adaptor protein Dab1 9-11 . Phosphorylated Dab1 is quickly degraded 1,2,5 . Thus, the amount of Dab1 is indicative of the strength of Reelin signaling [12][13][14][15] . Several pathways act downstream of Dab1 phosphorylation and regulate the cytoskeleton, adhesion molecules, ion channels, and neurotransmitter release 1-3,5,16 . Accordingly, insufficient Reelin signaling could contribute to the pathogenesis of neuropsychiatric disorders, such as epilepsy 17-19 , schizophrenia 6,16,20 , and autism 3,8,16 . Furthermore, recent studies showed that a decrease or insufficiency in Reelin signaling could aggravate Alzheimer's disease (AD) 13,21-26 . Therefore, it is important to understand the molecular mechanisms by which Reelin is regulated. Of particular importance is understanding how Reelin signaling is turned off because inhibition of such a mechanism would strengthen Reelin signaling and may help improve brain disorders caused by diminished Reelin function.Secreted signaling proteins can be inactivated or disabled by a variety of mechanisms, including internalization by its receptor, sequestration by an inactive (decoy) binding partner, and degradation. Internalization via ApoER2 and VLDLR 27,28 and a dominant-negative effect of the secreted extracellular domain of ApoER2 29 play a role in limiting Reelin signaling. Reelin protein (approximately 430 kDa as a monomer) contains three specific cleavage sites (N-t, C-t, and WC; reviewed in 7 ). N-t cleavage occurs between Pro1244 and Ala1245 to generate two fragments with molecular mas...

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“…Meprin α processes proMMP-9 and enhances its activation kinetics (Geurts et al, 2012). Additionally, meprin α is able to cleave proteins that are related to synaptic plasticity, such as Reelin (Sato et al, 2016) and vasoactive intestinal peptide (Sterchi et al, 2008). In addition, studies in non-mammalian species have demonstrated a role for Tll proteases in learning.…”

Section: Introductionmentioning

confidence: 99%

Multifaceted Roles of Metzincins in CNS Physiology and Pathology: From Synaptic Plasticity and Cognition to Neurodegenerative Disorders

Brzdąk

Nowak

Wiera

et al. 2017

Front. Cell. Neurosci.

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The extracellular matrix (ECM) and membrane proteolysis play a key role in structural and functional synaptic plasticity associated with development and learning. A growing body of evidence underscores the multifaceted role of members of the metzincin superfamily, including metalloproteinases (MMPs), A Disintegrin and Metalloproteinases (ADAMs), A Disintegrin and Metalloproteinase with Thrombospondin Motifs (ADAMTSs) and astacins in physiological and pathological processes in the central nervous system (CNS). The expression and activity of metzincins are strictly controlled at different levels (e.g., through the regulation of translation, limited activation in the extracellular space, the binding of endogenous inhibitors and interactions with other proteins). Thus, unsurprising is that the dysregulation of proteolytic activity, especially the greater expression and activation of metzincins, is associated with neurodegenerative disorders that are considered synaptopathies, especially Alzheimer’s disease (AD). We review current knowledge of the functions of metzincins in the development of AD, mainly the proteolytic processing of amyloid precursor protein, the degradation of amyloid β (Aβ) peptide and several pathways for Aβ clearance across brain barriers (i.e., blood-brain barrier (BBB) and blood-cerebrospinal fluid barrier (BCSFB)) that contain specific receptors that mediate the uptake of Aβ peptide. Controlling the proteolytic activity of metzincins in Aβ-induced pathological changes in AD patients’ brains may be a promising therapeutic strategy.

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Determination of cleavage site of Reelin between its sixth and seventh repeat and contribution of meprin metalloproteases to the cleavage

Cited by 26 publications

References 44 publications

Physiological significance of proteolytic processing of Reelin revealed by cleavage-resistant Reelin knock-in mice

Physiological significance of proteolytic processing of Reelin revealed by cleavage-resistant Reelin knock-in mice

Physiological significance of proteolytic processing of Reelin revealed by cleavage-resistant Reelin knock-in mice

Multifaceted Roles of Metzincins in CNS Physiology and Pathology: From Synaptic Plasticity and Cognition to Neurodegenerative Disorders

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