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Two‐dimensional electrophoresis of proteins in human retinal pigment epithelial cells: Identification of cytoskeletal proteins
Author(s) -
Haley James E.,
Flood Mary T.,
Kjeldbye Hild M.,
Maiello Eda M.,
Bilek Mary K.,
Gouras Peter
Publication year - 1983
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150040206
Subject(s) - isoelectric focusing , cytoskeleton , retinal pigment epithelium , gel electrophoresis , isoelectric point , keratin , pigment , retinal , actin , electrophoresis , two dimensional gel electrophoresis , biology , epithelium , biochemistry , chemistry , microbiology and biotechnology , cell , proteomics , enzyme , genetics , organic chemistry , gene , paleontology
Two‐dimensional gel (2‐D) electrophoresis coupled to fluorography was used to obtain the [ 35 S]‐methionine labeled protein profiles of cultured human retinal pigment epithelial cells. These patterns revealed about 250 polypeptides in the largely acidic isoelectric focusing 2‐D profile and approximately 200 in the mainly basic non‐equilibrium pH gradient electrophoresis 2‐D profile. The isolation of a detergent‐resistant cytoskeletal preparation revealed an enriched content of intermediate‐sized (7–10 nm) filaments as shown by electron microscopy. Two‐dimensional analysis (acidic isoelectric focusing, 2‐D) of the cytoskeletal preparation showed about 30 major proteins including non‐muscle actin, which had their counterparts in the total retinal pigment epithelium protein profile. These studies have begun to establish the subcellular local (filaments) and identity of some of the currently visualized 450 different retinal pigment epithelium proteins.