Ana-Maria Enciu
Carol Davila University of Medicine,Romania
Title: Electrophoretic analysis of amyloid precursor protein oligomers separated from mouse brain cortex
Biography
Biography: Ana-Maria Enciu
Abstract
Background: Amyloid precursor protein (APP) is a membrane protein highly expressed in the brain, involved in the pathogeny of Alzheimer`s disease. Recently, protein dimers were reported to occur in vitro, with impact on pathologic metabolization and senile plaque formation.
Aim: Our study investigated whether the expression of APP is dependent on a membrane scaffolding protein – caveolin and if in native state APP can be found in dimeric or oligomeric complexes.
Material and method: brain cortex was prelevated from 7-9 mo old normal and caveolin knockout mice and investigated for amyloid precursor protein by reducing and non-reducing electrophoresis, followed by western blotting. Membranes were next separated by ultracentrifugation and investigated for APP containing macromolecular complexes by non-denaturating, blue native electrophoresis
Results APP expression was modified by the absence of caveolin, depending on the brain region. Electrophoretic separation of macromolecular complexes from mouse brain cortex was dependant on the extraction buffer and type of gel. No dimeric or low-rank oligomers were highlited with the selected protocols.
Conclusion: APP dimers were not present in the investigated brain regions even under mild protein extraction conditions. We propose that dimerization is either an in vitro occurrence, or is masked by association with another membrane proteins in macromolecular complexes.
Acknowledgment: This work was supported by a grant of the Romanian National Authority for Scientific Research and Innovation CNCS-UEFISCDI, project number PN-II-RU-TE-2014-4-1534.