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Functional reassembly of the anion transport domain of human red cell band 3 (AE1) from multiple and non‐complementary fragments
Author(s) -
Groves Jonathan D.,
Wang Lin,
Tanner Michael J.A.
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00909-0
Subject(s) - band 3 , chemistry , red cell , domain (mathematical analysis) , biophysics , biochemistry , biology , membrane protein , computer science , artificial intelligence , membrane , mathematics , mathematical analysis
We constructed cDNA clones encoding N‐terminal, C‐terminal and internal polypeptide fragments of the human red cell anion exchanger (band 3; AE1). The internal fragments comprised between one and seven putative transmembrane spans with two or more spans deleted from both termini of the membrane domain of band 3. Sets of three, four or five complementary fragments, which together represented the complete amino acid sequence of the membrane domain, were co‐expressed in Xenopus oocytes. Stilbene disulphonate‐sensitive chloride uptake assays revealed that all six of the three‐fragment combinations and two of the four‐fragment combinations reassembled functionally in vivo. Unexpectedly, co‐expression of a non‐complementary pair of fragments comprising the first five and last seven putative transmembrane spans (i.e. entirely lacking spans six and seven) was also found to be sufficient to generate stilbene disulphonate‐sensitive chloride uptake.