title: Functional characterisation of ganglioside-induced differentiation-associated protein 1 as a glutathione transferase.
abstract: Mutations in the ganglioside-induced differentiation-associated protein 1 (GDAP1) gene have been linked with Charcot-Marie-Tooth (CMT) disease. This protein, and its paralogue GDAP1L1, appear to be structurally related to the cytosolic glutathione S-transferases (GST) including an N-terminal thioredoxin fold domain with conserved active site residues. The specific function, of GDAP1 remains unknown. To further characterise their structure and function we purified recombinant human GDAP1 and GDAP1L1 proteins using bacterial expression and immobilised metal affinity chromatography. Like other cytosolic GSTs, GDAP1 protein has a dimeric structure. Although the full-length proteins were largely insoluble, the deletion of a proposed C-terminal transmembrane domain allowed the preparation of soluble protein. The purified proteins were assayed for glutathione-dependent activity against a library of 'prototypic' GST substrates. No evidence of glutathione-dependent activity or an ability to bind glutathione immobilised on agarose was found.
authors.affiliation: Molecular Genetics Group, Division of Molecular Biosciences, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia.
authors.forename: Alison J
authors.initials: AJ
authors.lastname: Shield
authors.forename: Tracy P
authors.initials: TP
authors.lastname: Murray
authors.forename: Philip G
authors.initials: PG
authors.lastname: Board
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002460:Cell Line; D002607:Charcot-Marie-Tooth Disease / Q000235:genetics; D019281:Dimerization; D005982:Glutathione Transferase / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D016415:Sequence Alignment
chemical_list: C118663:GDAP protein; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase