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The N‐ and C‐termini of the tricarboxylate carrier are exposed to the cytoplasmic side of the inner mitochondrial membrane
Author(s) -
Capobianco L.,
Bisaccia F.,
Michel A.,
Sluse F.E.,
Palmieri F.
Publication year - 1995
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 - 10.1016/0014-5793(94)01379-f
Subject(s) - intermembrane space , polyclonal antibodies , cytoplasm , cytosol , inner membrane , transmembrane protein , chemistry , bacterial outer membrane , membrane , biochemistry , mitochondrial carrier , inner mitochondrial membrane , mitochondrion , microbiology and biotechnology , antibody , biophysics , biology , enzyme , receptor , escherichia coli , immunology , gene
Polyclonal antibodies were raised in rabbits against two synthetic peptides corresponding to the N‐ and C‐terminal regions of the rat‐liver mitochondria) tricarboxylate carrier. ELISA tests performed with intact and permeabilized rat‐liver mitoplasts showed that both anti‐N‐terminal and anti‐C‐terminal antibodies bind only to the cytoplasmic surface of the inner membrane, indicating that both termini of the membrane‐bound tricarboxylate carrier are exposed to the mitochondria) intermembrane space. Furthermore, tryptic digestion of intact mitoplasts markedly decreased the binding of anti‐N‐terminal and anti‐C‐terminal antibodies to the tricarboxylate carrier. These results are consistent with an arrangement of the tricarboxylate carrier monomer into an even number of transmembrane segments, with the N‐ and C‐termini protruding toward the cytosol.