title: Intracellular interactions between protein 4.1 and glycophorin C on transport vesicles, as determined by fluorescence correlation spectroscopy.
abstract: Interaction of protein 4.1 (4.1R) with the transmembrane protein glycophorin C (GPC) regulates the functions of erythrocyte membrane. Fluorescence correlation spectroscopy (FCS) was used to define the interaction of EGFP-4.1R with DsRed-GPC on transport vesicles (TVs) by measuring their fluctuation in living cells. DsRed-GPC expressed in HeLa cells was delivered to the plasma membrane through slow vesicle transport. EGFP-4.1R, which freely diffused in the cytosol when expressed alone, diffused slowly when co-expressed with DsRed-GPC, indicating association of EGFP-4.1R with TVs. Fluorescence cross-correlation spectroscopy (FCCS) showed direct interaction of EGFP-4.1R with DsRed-GPC on TVs. The present study demonstrates that 4.1R binds to GPC on TVs in living cells.
authors.affiliation: Department of Biochemistry, School of Medicine, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Shotaro
authors.initials: S
authors.lastname: Tanaka
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Takakuwa
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D006021:Glycophorins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence / Q000379:methods*; D022161:Transport Vesicles / Q000737:chemistry* / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D005456:Fluorescent Dyes; D006021:Glycophorins; D008565:Membrane Proteins; C485184:enhanced green fluorescent protein; C028841:erythrocyte membrane band 4.1 protein; D049452:Green Fluorescent Proteins