Two-dimensional polyacrylamide gel electrophoresis of water-soluble erythrocyte membrane proteins

✍ Scribed by Mark D. White; Gregory B. Ralston

• Publisher: John Wiley and Sons
• Year: 1981
• Tongue: English
• Weight: 758 KB
• Volume: 2
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150020408

✦ Synopsis

Abstract

A method is reported for analyzing the oligomeric state of proteins in a mixture by multidimensional electrophoresis. Water‐soluble proteins prepared from human erythrocyte membranes were separated by means of gel filtration followed by electrophoresis in detergent‐free disc gels. The major bands in detergent‐free disc gels were identified by means of electrophoresis into a second‐dimensional slab gel containing sodium dodecyl sulphate. Detergent‐free disc gels were also subjected to two‐dimensional electrophoresis into detergent‐free slab gels consisting of a continuous gradient of polyacrylamide gel. It was shown that many of the water‐soluble proteins existed as distinct oligomers. Spectrin aggregates (tetramer and dimmer) accounted for the slowest moving bands in the detergent‐free disc gels. A water‐soluble protein of the component 3 region appeared to be present as a hexamer, while component 4.1 was present as a tetramer. Components 4.3, 4.9 and 5 appeared to be present in a large number of aggregated states. Components 7 and 8 formed a heteropolymeric protein complex.