title: Isolation and cloning of a voltage-dependent anion channel-like Mr 36,000 polypeptide from mammalian brain.
abstract: A polypeptide of M(r) 36,000 (36 kDa) was isolated from detergent-solubilized membrane fractions of mammalian brain on a benzodiazepine affinity column utilized for the purification of the gamma-aminobutyric acid/benzodiazepine receptor protein, followed by preparative gel electrophoresis. Partial protein sequence for two fragments of the 36-kDa polypeptide allowed the isolation of cDNA clones from a rat hippocampal library. An open reading frame coding a sequence of 295 amino acid residues containing the two probe peptide sequences with minor differences, and a putative N-terminal signal peptide of 25 residues was found. Hydropathy index revealed no regions of alpha-helix suitable for membrane spanning, but several areas of alternating hydrophilic and hydrophobic residues consistent with beta-strands. The sequence of this brain protein was 24% identical to that of a yeast mitochondrial protein, the voltage-dependent anion channel (VDAC), and over 70% identical with the VDAC from human B lymphocytes. The gamma-aminobutyric acid type A (GABAA) receptor/36-kDa preparation purified on benzodiazepine affinity column has channel-forming activity in lipid bilayer membranes that is virtually identical to VDAC isolated from mitochondria of various sources, indicating that the 36-kDa protein is a new member of the VDAC family of proteins. An antiserum raised against the purified 36-kDa polypeptide was able to precipitate [3H]muscimol binding activity, indicating a tight association with the GABAA receptor protein in vitro and copurification on the benzodiazepine affinity column due to this association. Further studies are needed to determine whether such an association occurs in vivo.
authors.affiliation: Department of Pharmacology, School of Medicine, University of California, Los Angeles 90024.
authors.forename: M H
authors.initials: MH
authors.lastname: Bureau
authors.forename: M
authors.initials: M
authors.lastname: Khrestchatisky
authors.forename: M A
authors.initials: MA
authors.lastname: Heeren
authors.forename: E B
authors.initials: EB
authors.lastname: Zambrowicz
authors.forename: H
authors.initials: H
authors.lastname: Kim
authors.forename: T M
authors.initials: TM
authors.lastname: Grisar
authors.forename: M
authors.initials: M
authors.lastname: Colombini
authors.forename: A J
authors.initials: AJ
authors.lastname: Tobin
authors.forename: R W
authors.initials: RW
authors.lastname: Olsen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000838:Anions; D001483:Base Sequence; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007473:Ion Channels; D008969:Molecular Sequence Data; D051381:Rats; D011963:Receptors, GABA-A / Q000378:metabolism
chemical_list: D000838:Anions; D007473:Ion Channels; D011963:Receptors, GABA-A; D004247:DNA