Curation Details
Interaction ID: IM-22281-4

Unique identifier for interactor Auniprotkb:P56817
Unique identifier for interactor Buniprotkb:O75955
Alternative identifier for interactor Aintact:EBI-2433139
Alternative identifier for interactor Bintact:EBI-603643
Aliases for Apsi-mi:bace1_human(display_long)
uniprotkb:BACE(gene name synonym)
uniprotkb:KIAA1149(gene name synonym)
uniprotkb:Beta-site amyloid precursor protein cleaving enzyme 1(gene name synonym)
uniprotkb:Membrane-associated aspartic protease 2(gene name synonym)
uniprotkb:Memapsin-2(gene name synonym)
uniprotkb:Aspartyl protease 2(gene name synonym)
uniprotkb:BACE1(gene name)
Aliases for Bpsi-mi:flot1_human(display_long)
uniprotkb:FLOT1(gene name)
Interaction detection methodspsi-mi:"MI:0813"(proximity ligation assay)
First authorJohn et al. (2014)
Identifier of the publicationpubmed:24612608
NCBI Taxonomy identifier for interactor Ataxid:9606(human)
taxid:9606(Homo sapiens)
NCBI Taxonomy identifier for interactor Btaxid:9606(human)
taxid:9606(Homo sapiens)
Interaction typespsi-mi:"MI:2364"(proximity)
Source databases and identifierspsi-mi:"MI:0471"(MINT)
Interaction identifier(s) in the corresponding source databaseintact:EBI-9213821
Confidence scoreintact-miscore:0.27
Complex expansion-
Biological role AUnspecified role
Biological role BUnspecified role
Experimental role ANeutral component
Experimental role BNeutral component
Interactor type AProtein
Interactor type BProtein
Annotations for the interactionfigure legend:f2b
comment:"\"To dissect the role of flotillins in BACE1 binding in more detail and to test if the interaction is a direct one, we performed direct pulldown assays using bacterially expressed, purified proteins (Figure 1D). Purified flotillin-1 again strongly bound to the WT BACE1-GST tail, whereas no binding was observed with the LLAA tail. Only a very weak binding that was close to the GST background level was observed with purified flotillin-2. Taken together, these data show that flotillin-1 directly interacts with the cytoplasmic tail of BACE1 and that this interaction is mediated by the well known dileucine sorting motif in BACE1. To corroborate these findings in an in vivo context, we performed coimmunoprecipitation experiments with endogenous flotillins and BACE1 (Figure 2A). For this, we used polyclonal flotillin-2 antibodies since they result in a quantitative precipitation of the pool of flotillin-1/flotillin-2 complexes in the cells. BACE1 was found to be immunoprecipitated with endogenous flotillin-2, verifying the relevance of the pulldown data. Since the signals observed in the coprecipitation experiment were very weak, we used the Proximity Ligation Assay (PLA) to verify the interaction of endogenous BACE1 and flotillins. PLA is based on the use of specific antibodies and can detect two proteins that are at very close vicinity (less than 40 nm) in cells and is generally used to study interaction of proteins [38-40]. Interacting proteins are visualized as fluorescent signals whose localization in the cells can also be assessed, giving information on the compartment in which the interaction takes place. However, the signal intensities as such do not correlate with the strength or quantity of the interaction. Quantification of the signals is done by counting the amount of fluorescent signals per cells and comparing these to negative controls (performed without the first antibody). PLA analysis of endogenous flotillin-1 or flotillin-2 and BACE1 showed that the fluorescent dots that arise upon interaction of these proteins were mainly detected in the perinuclear region of the cells, with few dots residing in the cell periphery (Fig. 2B).\""
dataset:Alzheimers - Interactions investigated in the context of Alzheimers disease
full coverage:Only protein-protein interactions
curation depth:imex curation
NCBI Taxonomy identifier for the host organismtaxid:9606(human-hela)
taxid:9606(Homo sapiens HeLa epitheloid cervical carcinoma cell)
Parameters of the interaction-
Creation date2014/02/21
Update date2017/06/28
negative Boolean valuefalse
Feature(s) for interactor A-
Feature(s) for interactor B-
Stoichiometry for interactor A-
Stoichiometry for interactor B-
Participant identification method for interactor AImmunostaining
Participant identification method for interactor BImmunostaining