Isolation and characterization of the rainbow trout erythrocyte band-3 protein.
Rainbow trout (Salmo gairdneri) band-3 protein was isolated from trout erythrocyte plasma membranes by a combination of preparative SDS/PAGE and electroelution. High purity and recovery of the plasma membranes were achieved by a new method. This was demonstrated using 4,4'diiso-thiocyano[3H2]dihydro-stilbene 2,2'disulfonic acid (3H2DIDS) which specifically labels band-3 protein. On SDS/PAGE, band-3 protein yields a similarly diffuse pattern, as does mammalian band-3 protein, with an apparent Mr of 116,000. In situ chymotryptic cleavage/cross-linking experiments with 3H2DIDS reveal that the fragments cross-link as in human and mouse band-3 proteins but that there are minor differences. Treatment of trout erythrocytes with trypsin results in cleavage of the band-3 protein. Purified polyclonal antibodies raised against trout band-3 protein react with trout band-3 protein and do not crossreact with mouse or human band-3 protein. They react specifically with only one chymotryptic fragment of trout band-3 protein.
Michel
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F
Max-Planck-Institut für Biophysik, Abteilung Zellphysiologie, Frankfurt/Main, FRG.
Rudloff
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V
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000302:isolation & purification* / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism; D008970:Molecular Weight; D015388:Organelles / Q000378:metabolism; D011485:Protein Binding; D012487:Salmonidae / Q000097:blood*; D014337:Trout / Q000097:blood*; D014357:Trypsin
D001457:Anion Exchange Protein 1, Erythrocyte; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D014357:Trypsin; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid