Purification of erythrocyte protein 4.1 by selective interaction with inositol hexaphosphate.
Protein 4.1 is a multifunctional structural protein occupying a strategic position in the erythrocyte membrane. It is present in the erythrocyte membrane skeleton and in many nonerythroid cells. This report describes a novel method for purifying this protein based on its selective interaction with inositol hexaphosphate dimagnesium tetrapotassium salt. This interaction was discovered in the course of chromatography of high-salt extract of inside-out membrane vesicles on Procion orange MX-2R-Sepharose. The new procedure is simple and selective and produces protein 4.1 with better yield than that obtained with a previously published procedure. The purified protein 4.1 has the same immunoreactivity and the same alpha-chymotryptic digest profile as protein 4.1 purified by published methods and is fully functional in enhancing the interaction between F-actin and spectrin dimers.
el Ouggouti
S
S
INSERM U 160, Hôpital Beaujon, Clichy, France.
Bournier
O
O
Boivin
P
P
Bertrand
O
O
Dhermy
D
D
D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D002918:Chymotrypsin / Q000378:metabolism; D003598:Cytoskeletal Proteins; D004910:Erythrocyte Membrane / Q000737:chemistry*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D009479:Neuropeptides; D010446:Peptide Fragments / Q000302:isolation & purification; D010833:Phytic Acid / Q000378:metabolism*; D012685:Sepharose; D012995:Solubility; D014227:Triazines
D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D009479:Neuropeptides; D010446:Peptide Fragments; D014227:Triazines; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; C041397:Procion Orange MX-2R; D010833:Phytic Acid; D012685:Sepharose; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D002918:Chymotrypsin