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Título

Heteromers of amyloid precursor protein in cerebrospinal fluid

AutorCuchillo-Ibáñez, Inmaculada CSIC ORCID; López-Font, Inmaculada CSIC ORCID; Boix-Amorós, Alba CSIC ORCID; Brinkmalm, Gunnar; Blennow, Kaj; Molinuevo, José Luis; Sáez-Valero, Javier CSIC ORCID
Palabras clavesAPPα
sAPPβ
Heteromers
Cerebrospinal fluid
Alzheimer’s disease
ELISA
Fecha de publicación8-ene-2015
EditorBioMed Central
CitaciónMolecular Neurodegeneration 10(2): 1-11(2015)
ResumenBackground: Soluble fragments of the amyloid precursor protein (APP) generated by α- and β-secretases, sAPPα and sAPPβ, have been postulated as promising new cerebrospinal fluid (CSF) biomarkers for the clinical diagnosis of Alzheimer’s disease (AD). However, the capacity of these soluble proteins to assemble has not been explored and could be relevant. Our aim is to characterize possible sAPP oligomers that could contribute to the quantification of sAPPα and sAPPβ in CSF by ELISA, as well as to characterize the possible presence of soluble full-length APP (sAPPf).
Results: We employed co-immunoprecipitation, native polyacrylamide gel electrophoresis and ultracentrifugation in sucrose density gradients to characterize sAPP oligomers in CSF. We have characterized the presence of sAPPf in CSF from NDC and AD subjects and demonstrated that all forms, including sAPPα and sAPPβ, are capable of assembling into heteromers, which differ from brain APP membrane-dimers. We measured sAPPf, sAPPα and sAPPβ by ELISA in CSF samples from AD (n = 13) and non-disease subjects (NDC, n = 13) before and after immunoprecipitation with antibodies against the C-terminal APP or against sAPPβ. We demonstrated that these sAPP heteromers participate in the quantification of sAPPα and sAPPβ by ELISA. Immunoprecipitation with a C-terminal antibody to remove sAPPf reduced by ~30% the determinations of sAPPα and sAPPβ by ELISA, whereas immunoprecipitation with an APPβ antibody reduced by ~80% the determination of sAPPf and sAPPα.
Conclusions: The presence of sAPPf and sAPP heteromers should be taken into consideration when exploring the levels of sAPPα and sAPPβ as potential CSF biomarkers.
Versión del editorhttp://dx.doi.org/10.1186/1750-1326-10-2
URIhttp://hdl.handle.net/10261/122961
DOI10.1186/1750-1326-10-2
ISSN1750-1326
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