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Full-Text Articles in Life Sciences
Detecting Mitochondrial Permeability Transition By Confocal Imaging Of Intact Cells Pinocytically Loaded With Calcein, Rachel Jones, Alison Smail, Mark Wilson
Detecting Mitochondrial Permeability Transition By Confocal Imaging Of Intact Cells Pinocytically Loaded With Calcein, Rachel Jones, Alison Smail, Mark Wilson
Mark R Wilson
When studied in vitro, mitochondrial permeability transition (MPT) is associated with an increase in mitochondrial permeability to solutes up to 1500 Da in mass and a loss of electrical potential difference across the inner mitochondrial membrane (delta psi mit). The MPT has been implicated as being important in cellular calcium homeostasis, autophagy and cell death via necrosis and apoptosis. Thus, it is important to develop a valid technique for accurate measurement of this phenomenon in intact cells. We developed a procedure for the detection of MPT in intact cells that avoids disadvantages associated with earlier approaches. In this new technique, …
Evidence That Clusterin Has Discrete Chaperone And Ligand Binding Sites, Johnathon Lakins, Stephen Poon, Simon Easterbrook-Smith, John Carver, Martin Tenniswood, Mark Wilson
Evidence That Clusterin Has Discrete Chaperone And Ligand Binding Sites, Johnathon Lakins, Stephen Poon, Simon Easterbrook-Smith, John Carver, Martin Tenniswood, Mark Wilson
Mark R Wilson
Clusterin is the first identified extracellular mammalian chaperone and binds to a wide variety of partly unfolded, stressed proteins. Clusterin also binds to many different unstressed ligands including the cell surface receptor low density lipoprotein receptor-related protein-2 (LRP-2). It is unknown whether clusterin binds to all of these many ligands via one or more binding sites. Furthermore, the region(s) of clusterin involved in these many binding interactions remain to be identified. As part of an investigation of these issues, we expressed recombinant human clusterin in the yeast Pichia pastoris. The resultant protein had variable proteolytic truncations of the C-terminal region …