A surface plasmon resonance-based biosensor with full-length BACE1 in a reconstituted membrane

Stenberg, Gun (författare): Uppsala universitet,Institutionen för biokemi och organisk kemi

Gossas, Thomas (författare): Uppsala universitet,Institutionen för biokemi och organisk kemi

Nyström, Susanne (författare)

Baraznenok, Vera (författare)

Lindström, Erik (författare)

Danielson, U. Helena, 1959- (författare): Uppsala universitet,Biokemi,U Helena Danielson

(creator_code:org_t)

Elsevier BV, 2011
2011
Engelska.
Ingår i: Analytical Biochemistry. - : Elsevier BV. - 0003-2697 .- 1096-0309. ; 414:1, s. 14-22

Relaterad länk:: https://urn.kb.se/re...; visa fler...; https://doi.org/10.1...; visa färre...

Abstract Ämnesord

Stäng

A surface plasmon resonance (SPR) biosensor-based assay for membrane-embedded full-length BACE1 (β-site amyloid precursor protein cleaving enzyme 1), a drug target for Alzheimer's disease, has been developed. It allows the analysis of interactions with the protein in its natural lipid membrane environment. The enzyme was captured via an antibody recognizing a C-terminal His6 tag, after which a lipid membrane was reconstituted on the chip using a brain lipid extract. The interaction between the enzyme and several inhibitors confirmed that the surface was functional. It had slightly different interaction characteristics as compared with a reference surface with immobilized ectodomain BACE1 but had the same inhibitor characteristic pH effect. The possibility of studying interactions with BACE1 under more physiological conditions than assays using truncated enzyme or conditions dictated by high enzyme activity is expected to increase our understanding of the role of BACE1 in Alzheimer's disease and contribute to the discovery of clinically efficient BACE1 inhibitors. The strategy exploited in the current study can be adapted to other membrane-bound drug targets by selecting suitable capture antibodies and lipid mixtures for membrane reconstitution.

Av författaren/redakt...: Christopeit, Ton ...; Stenberg, Gun; Gossas, Thomas; Nyström, Susanne; Baraznenok, Vera; Lindström, Erik; visa fler...; Danielson, U. He ...; visa färre...

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