title: Disassembly of the cytosolic chaperonin in mammalian cell extracts at intracellular levels of K+ and ATP.
abstract: The eukaryotic, cytoplasmic chaperonin, CCT, is essential for the biogenesis of actin- and tubulin-based cytoskeletal structures. CCT purifies as a doubly toroidal particle containing two eight-membered rings of approximately 60-kDa ATPase subunits, each encoded by an essential and highly conserved gene. However, immunofluorescence detection with subunit-specific antibodies has indicated that in cells CCT subunits do not always co-localize. We report here that CCT ATPase activity is highly dependent on K+ ion concentration and that in cell extracts, at physiological levels of K+ and ATP, there is considerable dissociation of CCT to a smaller oligomeric structure and free subunits. This dissociation is consequent to ATP hydrolysis and is readily reversed on removal of ATP. The ranking order for ease with which subunits can exit the chaperonin particle correlates well with the length of a loop structure, identified by homology modeling, in the intermediate domain of CCT subunits. K+-ATP-induced disassembly is not an intrinsic property of purified CCT over a 40-fold concentration range and requires the presence of additional factor(s) present in cell extracts.
authors.affiliation: Department of Biosciences, University of Kent, Canterbury CT2 7NJ, United Kingdom. a.roobol@ukc.ac.uk
authors.forename: A
authors.initials: A
authors.lastname: Roobol
authors.forename: J
authors.initials: J
authors.lastname: Grantham
authors.forename: H C
authors.initials: HC
authors.lastname: Whitaker
authors.forename: M J
authors.initials: MJ
authors.lastname: Carden
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins / Q000378:metabolism*; D002850:Chromatography, Gel; D051379:Mice; D008958:Models, Molecular; D011188:Potassium / Q000378:metabolism*; D011487:Protein Conformation; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
chemical_list: C533315:Tcp1 protein, mouse; D000255:Adenosine Triphosphate; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins; D011188:Potassium