Partial characterization of the cytoplasmic domain of human kidney band 3.
The major anion exchanger in type A intercalated cells of the cortical and medullary collecting ducts of the human kidney is a truncated isoform of erythrocyte band 3 (AE1) that lacks the N-terminal 65 residues. Because this missing sequence has been implicated in the binding of ankyrin, protein 4.1, several glycolytic enzymes, hemoglobin, and hemichromes in erythrocytes, we have undertaken examination of the structure and peripheral protein interactions of this kidney isoform. The cytoplasmic domain of kidney band 3, kidney CDB3, was expressed in Escherichia coli and purified to homogeneity. The kidney isoform exhibited a circular dichroism spectrum and Stokes radius similar to its larger erythrocyte counterpart. Kidney CDB3 was also observed to engage in the same conformational equilibrium characteristic of erythrocyte CDB3. In contrast, the tryptophan and cysteine clusters of kidney CDB3 behaved very differently from erythrocyte CDB3 in response to pH changes and oxidizing conditions. Furthermore, kidney CDB3 did not bind ankyrin, protein 4.1, or aldolase, and expression of erythrocyte CDB3 was toxic to its bacterial host, whereas expression of kidney CDB3 was not. Taken together, these data suggest that the absence of the N-terminal 65 amino acids in kidney CDB3 eliminates the major function currently ascribed to CDB3 in erythrocytes, i.e. that of peripheral protein binding. The primary function of residues 66-379 found in kidney CDB3 thus remains to be elucidated.
Wang
C C
CC
Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, USA.
Moriyama
R
R
Lombardo
C R
CR
Low
P S
PS
D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D001483:Base Sequence; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D017931:DNA Primers; D004912:Erythrocytes / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation
D001457:Anion Exchange Protein 1, Erythrocyte; D017931:DNA Primers