Preparative isolation of glycoproteins from plasma membranes of different rat organs.
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.
Josić
D
D
Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, Dahlem, F.R.G.
Zeilinger
K
K
Lim
Y P
YP
Raps
M
M
Hofmann
W
W
Reutter
W
W
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D013056:Spectrophotometry, Ultraviolet
D008562:Membrane Glycoproteins; D008565:Membrane Proteins