N-glycosylation of human nicastrin is required for interaction with the lectins from the secretory pathway calnexin and ERGIC-53

Brito, Catarina; Pijak, Donald S.; Crystal, Adam S.; Fortna, Ryan R.; Li, Tong; Wong, Phil; Doms, Robert W.; Costa, Júlia

doi:10.1016/j.bbadis.2006.06.018

Cited by 20 publications

(15 citation statements)

References 40 publications

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“…We confirmed the association of CNX and NCT as previously described (Fig. 8A) (45). Moreover, this interaction was significantly reduced by the overexpression of 5201F (Fig.…”

Section: Binding Of 5201f Prevents the Glycosyl And The Structural Masupporting

confidence: 78%

Single Chain Variable Fragment against Nicastrin Inhibits the γ-Secretase Activity

Hayashi

Takatori

Urano

et al. 2009

Journal of Biological Chemistry

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␥-Secretase is a membrane protein complex that catalyzes intramembrane proteolysis of a variety of substrates including the amyloid ␤ precursor protein of Alzheimer disease. Nicastrin (NCT), a single-pass membrane glycoprotein that harbors a large extracellular domain, is an essential component of the ␥-secretase complex. Here we report that overexpression of a single chain variable fragment (scFv) against NCT as an intrabody suppressed the ␥-secretase activity. Biochemical analyses revealed that the scFv disrupted the proper folding and the appropriate glycosyl maturation of the endogenous NCT, which are required for the stability of the ␥-secretase complex and the intrinsic proteolytic activity, respectively, implicating the dual role of NCT in the ␥-secretase complex. Our results also highlight the importance of the calnexin cycle in the functional maturation of the ␥-secretase complex. The engineered intrabodies may serve as rationally designed, molecular targeting tools for the discovery of novel actions of the membrane proteins.

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“…We confirmed the association of CNX and NCT as previously described (Fig. 8A) (45). Moreover, this interaction was significantly reduced by the overexpression of 5201F (Fig.…”

Section: Binding Of 5201f Prevents the Glycosyl And The Structural Masupporting

confidence: 78%

Single Chain Variable Fragment against Nicastrin Inhibits the γ-Secretase Activity

Hayashi

Takatori

Urano

et al. 2009

Journal of Biological Chemistry

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“…In addition to FV and FVIII, other potential LMAN1 cargo proteins have been identified. [26][27][28][29][30][31] Some of these cargo proteins, such as CatC and CatZ, do not seem to require MCFD2. 24 Thus, MCFD2 may be a specific cargo selection protein for the recruitment of FV and FVIII, but not other cargo proteins, such as CatC and CatZ.…”

Section: Discussionmentioning

confidence: 99%

EF-hand domains of MCFD2 mediate interactions with both LMAN1 and coagulation factor V or VIII

Zheng

Liu

Zhou

et al. 2010

Blood

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Combined deficiency of factor V and factor VIII (F5F8D) is a bleeding disorder caused by mutations in either LMAN1 or MCFD2. LMAN1 (ERGIC-53) and MCFD2 form a Ca 2؉ -dependent cargo receptor that cycles between the endoplasmic reticulum (ER) and the ER-Golgi intermediate compartment for efficient transport of FV/FVIII from the ER to the Golgi. Here we show that the C-terminal EF-hand domains are both necessary and sufficient for MCFD2 to interact with LMAN1. MCFD2 with a deletion of the entire N-terminal non-EF hand region still retains the LMAN1-binding function. Deletions that disrupt core structure of the EF-hand domains abolish LMAN1 binding. Circular dichroism spectroscopy studies on missense mutations localized to different structural elements of the EF-hand domains suggest that Ca 2؉ -induced folding is important for LMAN1 interaction. The EF-hand domains also mediate the interaction with FV and FVIII. However, mutations in MCFD2 that disrupt the tertiary structure and abolish LMAN1 binding still retain the FV/FVIII binding activities, suggesting that this interaction is independent of Ca 2؉ -induced folding of the protein. Our results suggest that the EF-hand domains of MCFD2 contain separate binding sites for LMAN1 and FV/FVIII that are essential for cargo receptor formation and cargo loading in the ER. (Blood.

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“…Thus, ERGIC-53, like sortilin, may serve a transport function. Nicastrin, a glycoprotein component of the ␥-secretase complex, interacts with ERGIC-53 (113). However, a role of ERGIC-53 in AD has not been established.…”

Section: Discussionmentioning

confidence: 99%

Protection against β-Amyloid-induced Apoptosis by Peptides Interacting with β-Amyloid

Nelson

Alkon

2007

Journal of Biological Chemistry

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␤-Amyloid peptide produces apoptosis in neurons at micromolar concentrations, but the mechanism by which ␤-amyloid exerts its toxic effect is unknown. The normal biological function of ␤-amyloid is also unknown. We used phage display, coprecipitation, and mass spectrometry to examine the proteinprotein interactions of ␤-amyloid in normal rabbit brain in order to identify the biochemical receptors for ␤-amyloid. ␤-Amyloid was found to bind primarily to proteins involved in low density lipoprotein and cholesterol transport and metabolism, including sortilin, endoplasmic reticulum-Golgi intermediate compartment 2 (ERGIC2), ERGIC-53, steroid 5␣-reductase, and apolipoprotein B. ␤-Amyloid also bound to the C-reactive protein precursor, a protein involved in inflammation, and to 14-3-3, a protein that regulates glycogen synthase kinase-3␤, the kinase involved in tau phosphorylation. Of eight synthetic peptides identified as targets of ␤-amyloid, three were found to be effective blockers of the toxic effect of ␤-amyloid on cultured neuronal cells. These peptides bound to the hydrophobic region (residues 17-21) or to the nearby protein kinase C pseudo-phosphorylation site (residues 26 -30) of ␤-amyloid, suggesting that these may be the most critical regions for ␤-amyloid effector action and for aggregation. Peptides or other small molecules that bind to this region may protect against ␤-amyloid toxic effect by competitively blocking its ability to bind ␤-amyloid effector proteins such as sortilin and 14-3-3.

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N-glycosylation of human nicastrin is required for interaction with the lectins from the secretory pathway calnexin and ERGIC-53

Cited by 20 publications

References 40 publications

Single Chain Variable Fragment against Nicastrin Inhibits the γ-Secretase Activity

Single Chain Variable Fragment against Nicastrin Inhibits the γ-Secretase Activity

EF-hand domains of MCFD2 mediate interactions with both LMAN1 and coagulation factor V or VIII

Protection against β-Amyloid-induced Apoptosis by Peptides Interacting with β-Amyloid

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