title: A structural basis for cellular uptake of GST-fold proteins.
abstract: It has recently emerged that glutathione transferase enzymes (GSTs) and other structurally related molecules can be translocated from the external medium into many different cell types. In this study we aim to explore in detail, the structural features that govern cell translocation and by dissecting the human GST enzyme GSTM2-2 we quantatively demonstrate that the α-helical C-terminal domain (GST-C) is responsible for this property. Attempts to further examine the constituent helices within GST-C resulted in a reduction in cell translocation efficiency, indicating that the intrinsic GST-C domain structure is necessary for maximal cell translocation capacity. In particular, it was noted that the α-6 helix of GST-C plays a stabilising role in the fold of this domain. By destabilising the conformation of GST-C, an increase in cell translocation efficiency of up to ∼2-fold was observed. The structural stability profiles of these protein constructs have been investigated by circular dichroism and differential scanning fluorimetry measurements and found to impact upon their cell translocation efficiency. These experiments suggest that the globular, helical domain in the 'GST-fold' structural motif plays a role in influencing cellular uptake, and that changes that affect the conformational stability of GST-C can significantly influence cell translocation efficiency.
authors: The John Curtin School of Medical Research, Australian National University, Canberra, Australian Capital Territory, Australia.
authors: Melanie J
authors: MJ
authors: Morris
authors: Dan
authors: D
authors: Liu
authors: Llara M
authors: LM
authors: Weaver
authors: Philip G
authors: PG
authors: Board
authors: Marco G
authors: MG
authors: Casarotto
mesh_terms: D000818:Animals; D002460:Cell Line; D002942:Circular Dichroism; D005434:Flow Cytometry; D005470:Fluorometry; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D017433:Protein Structure, Secondary; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D005982:Glutathione Transferase; C412602:glutathione S-transferase Mu 2