title: The minimal structure containing the band 3 anion transport site. A 35Cl NMR study.
abstract: 35Cl NMR, which enables observation of chloride binding to the anion transport site on band 3, is used in the present study to determine the minimal structure containing the intact transport site. Removal of cytoskeletal and other nonintegral membrane proteins, or removal of the 40-kDa cytoskeletal domain of band 3, each leave the transport site intact. Similarly, cleavage of the 52-kDa transport domain into 17- and 35-kDa fragments by chymotrypsin leaves the transport site intact. Extensive proteolysis by papain reduces the integral red cell membrane proteins to their transmembrane segments. Papain treatment removes approximately 60% of the extramembrane portion of the transport domain and produces small fragments primarily in the range 3-7 kDa, with 5 kDa being most predominant. Papain treatment damages, but does not destroy, chloride binding to the transport site; thus, the minimal structure containing the transport site is composed solely of transmembrane segments. In short, the results are completely consistent with a picture in which the transport site is buried in the membrane where it is protected from proteolysis; the transmembrane segments that surround the transport site are held together by strong attractive forces within the bilayer; and the transport site is accessed by solution chloride via an anion channel leading from the transport site to the solution.
mesh_terms: D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000031:analogs & derivatives*; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives / Q000097:blood; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D000838:Anions; D001639:Bicarbonates / Q000097:blood; D001665:Binding Sites / Q000187:drug effects; D001692:Biological Transport; D002712:Chlorides / Q000097:blood*; D002918:Chymotrypsin / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000097:blood; D009994:Osmolar Concentration; D010206:Papain / Q000494:pharmacology; D010446:Peptide Fragments / Q000097:blood; D010658:Phenylglyoxal / Q000494:pharmacology; D013267:Stilbenes / Q000494:pharmacology
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D000838:Anions; D001639:Bicarbonates; D002712:Chlorides; D008565:Membrane Proteins; D010446:Peptide Fragments; D013267:Stilbenes; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; C018564:dihydro-DIDS; C022306:4,4'-dinitro-2,2'-stilbenedisulfonic acid; D002918:Chymotrypsin; D010206:Papain; D010658:Phenylglyoxal; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid