naa a22 4500 510746500 CHVBK 20180411083048.0 cr unu---uuuuu 180411e20130301xx s 000 0 eng 10.1007/s12192-012-0357-z doi (NATIONALLICENCE)springer-10.1007/s12192-012-0357-z Human TRiC complex purified from HeLa cells contains all eight CCT subunits and is active in vitro [Elektronische Daten] [Kelly Knee, Oksana Sergeeva, Jonathan King] Archaeal and eukaryotic cytosols contain group II chaperonins, which have a double-barrel structure and fold proteins inside a cavity in an ATP-dependent manner. The most complex of the chaperonins, the eukaryotic TCP-1 ring complex (TRiC), has eight different subunits, chaperone containing TCP-1 (CCT1-8), that are arranged so that there is one of each subunit per ring. Aspects of the structure and function of the bovine and yeast TRiC have been characterized, but studies of human TRiC have been limited. We have isolated and purified endogenous human TRiC from HeLa suspension cells. This purified human TRiC contained all eight CCT subunits organized into double-barrel rings, consistent with what has been found for bovine and yeast TRiC. The purified human TRiC is active as demonstrated by the luciferase refolding assay. As a more stringent test, the ability of human TRiC to suppress the aggregation of human γD-crystallin was examined. In addition to suppressing off-pathway aggregation, TRiC was able to assist the refolding of the crystallin molecules, an activity not found with the lens chaperone, α-crystallin. Additionally, we show that human TRiC from HeLa cell lysate is associated with the heat shock protein 70 and heat shock protein 90 chaperones. Purification of human endogenous TRiC from HeLa cells will enable further characterization of this key chaperonin, required for the reproduction of all human cells. Cell Stress Society International, 2012 TRiC nationallicence CCT nationallicence Chaperonin nationallicence Crystallin nationallicence Protein folding nationallicence Knee Kelly Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave., 68-330, 02139, Cambridge, MA, USA aut Sergeeva Oksana Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave., 68-330, 02139, Cambridge, MA, USA aut King Jonathan Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave., 68-330, 02139, Cambridge, MA, USA aut Cell Stress and Chaperones Springer Netherlands 18/2(2013-03-01), 137-144 1355-8145 18:2<137 2013 18 12192 https://doi.org/10.1007/s12192-012-0357-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12192-012-0357-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Knee Kelly Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave., 68-330, 02139, Cambridge, MA, USA aut NATIONALLICENCE 700 1- Sergeeva Oksana Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave., 68-330, 02139, Cambridge, MA, USA aut NATIONALLICENCE 700 1- King Jonathan Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave., 68-330, 02139, Cambridge, MA, USA aut NATIONALLICENCE 773 0- Cell Stress and Chaperones Springer Netherlands 18/2(2013-03-01), 137-144 1355-8145 18:2<137 2013 18 12192 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer