Preparative isolation of glycoproteins from plasma membranes of different rat organs

J Chromatogr. 1989 Dec 22:484:327-35. doi: 10.1016/s0021-9673(01)88980-9.

Abstract

By a combination of high-performance affinity chromatographic (HPAC) methods, several membrane proteins from liver, Morris hepatoma and kidney were isolated. The use of a tandem system, consisting of a concanavalin A (ConA) and a wheat germ agglutinin (WGA) high-performance liquid chromatographic (HPLC) column, as a first purification step allowed the isolation of proteins directly from organ homogenates. In a subsequent step, the membrane proteins can be isolated by simply using a combination of immunoaffinity HPLC and preparative sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). However, with these methods most proteins lose their biological activity. If native proteins are required, a combination of different HPAC methods has to be applied. Several membrane proteins were isolated in milligram amounts under non-denaturing conditions using either HPAC columns or Mem Sep membranes with immobilized lectins, collagen, amino acids, crown ethers or heparin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / metabolism
  • Chromatography, Affinity
  • Chromatography, High Pressure Liquid
  • Electrophoresis, Polyacrylamide Gel
  • Kidney / metabolism
  • Liver / metabolism
  • Liver Neoplasms, Experimental / metabolism
  • Male
  • Membrane Glycoproteins / isolation & purification*
  • Membrane Proteins / isolation & purification
  • Rats
  • Rats, Inbred Strains
  • Spectrophotometry, Ultraviolet

Substances

  • Membrane Glycoproteins
  • Membrane Proteins