Fluorescent labelling of membrane fatty acid transporter CD36 (SR-B2) in the extracellular loop.

ABSTRACT:

Context

Upon palmitate oversupply, membrane fatty acid-transporter CD36 (SR-B2) permanently translocates from endosomal storage to the sarcolemma, inducing lipotoxicity. CD36 translocation results from endosomal alkalinisation elicited by palmitate-induced disattachment of the cytoplasmic V1-subcomplex from the membrane-integrated V0-subcomplex of vacuolar-type H+-ATPase.

Objective

Develop a CD36 fluorescent labeling technique as initial step towards live cell imaging.

Methods

Three human CD36 (hCD36) mutants were constructed via insertion of a tetracysteine motif at different positions within the extracellular domain. Constructs were lentivirally transduced for subsequent CD36 labeling with fluorescein-arsenical hairpin-binder (FlAsH). Cell imaging was combined with

SUBMITTER: Liu Y 

PROVIDER: S-EPMC6343965 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature