PMID- 24457959
OWN - NLM
STAT- MEDLINE
DCOM- 20140902
LR  - 20211021
IS  - 2041-4889 (Electronic)
VI  - 5
IP  - 1
DP  - 2014 Jan 23
TI  - The fine-tuning of TRAF2-GSTP1-1 interaction: effect of ligand binding and in 
      situ detection of the complex.
PG  - e1015
LID - 10.1038/cddis.2013.529 [doi]
AB  - We provide the first biochemical evidence of a direct interaction between the 
      glutathione transferase P1-1 (GSTP1-1) and the TRAF domain of TNF 
      receptor-associated factor 2 (TRAF2), and describe how ligand binding modulates 
      such an equilibrium. The dissociation constant of the heterocomplex is K(d)=0.3 
      muM; however the binding affinity strongly decreases when the active site of 
      GSTP1-1 is occupied by the substrate GSH (K(d)>/=2.6 muM) or is inactivated by 
      oxidation (Kd=1.7 muM). This indicates that GSTP1-1's TRAF2-binding region 
      involves the GSH-binding site. The GSTP1-1 inhibitor NBDHEX further decreases the 
      complex's binding affinity, as compared with when GSH is the only ligand; this 
      suggests that the hydrophobic portion of the GSTP1-1 active site also contributes 
      to the interaction. We therefore hypothesize that TRAF2 binding inactivates 
      GSTP1-1; however, analysis of the data, using a model taking into account the 
      dimeric nature of GSTP1-1, suggests that GSTP1-1 engages only one subunit in the 
      complex, whereas the second subunit maintains the catalytic activity or binds to 
      other proteins. We also analyzed GSTP1-1's association with TRAF2 at the cellular 
      level. The TRAF2-GSTP1-1 complex was constitutively present in U-2OS cells, but 
      strongly decreased in S, G2 and M phases. Thus the interaction appears regulated 
      in a cell cycle-dependent manner. The variations in the levels of individual 
      proteins seem too limited to explain the complex's drastic decline observed in 
      cells progressing from the G0/G1 to the S-G2-M phases. Moreover, GSH's 
      intracellular content was so high that it always saturated GSTP1-1. 
      Interestingly, the addition of NBDHEX maintains the TRAF2-GSTP1-1 complex at low 
      levels, thus causing a prolonged cell cycle arrest in the G2/M phase. Overall, 
      these findings suggest that a reversible sequestration of TRAF2 into the complex 
      may be crucial for cell cycle progression and that multiple factors are involved 
      in the fine-tuning of this interaction.
FAU - De Luca, A
AU  - De Luca A
AD  - The NAST Centre for Nanoscience & Nanotechnology & Innovative Instrumentation, 
      University of Tor Vergata, Rome, Italy.
FAU - Mei, G
AU  - Mei G
AD  - Department of Experimental Medicine and Surgery, University of Tor Vergata, Rome, 
      Italy.
FAU - Rosato, N
AU  - Rosato N
AD  - Department of Experimental Medicine and Surgery, University of Tor Vergata, Rome, 
      Italy.
FAU - Nicolai, E
AU  - Nicolai E
AD  - Department of Experimental Medicine and Surgery, University of Tor Vergata, Rome, 
      Italy.
FAU - Federici, L
AU  - Federici L
AD  - Department of Biomedical Sciences, University of Chieti G D'Annunzio, CeSI Center 
      of Excellence on Aging, Chieti, Italy.
FAU - Palumbo, C
AU  - Palumbo C
AD  - Department of Clinical Sciences and Translational Medicine, University of Tor 
      Vergata, Rome, Italy.
FAU - Pastore, A
AU  - Pastore A
AD  - Laboratory of Metabolomics and Proteomics, Children's Hospital IRCCS Bambino 
      Gesu, Rome, Italy.
FAU - Serra, M
AU  - Serra M
AD  - Laboratory of Experimental Oncology, Istituto Ortopedico Rizzoli IRCCS, Bologna, 
      Italy.
FAU - Caccuri, A M
AU  - Caccuri AM
AD  - 1] The NAST Centre for Nanoscience & Nanotechnology & Innovative Instrumentation, 
      University of Tor Vergata, Rome, Italy [2] Department of Experimental Medicine 
      and Surgery, University of Tor Vergata, Rome, Italy.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140123
PL  - England
TA  - Cell Death Dis
JT  - Cell death & disease
JID - 101524092
RN  - 0 (Ligands)
RN  - 0 (TNF Receptor-Associated Factor 2)
RN  - EC 2.5.1.18 (GSTP1 protein, human)
RN  - EC 2.5.1.18 (Glutathione S-Transferase pi)
RN  - GAN16C9B8O (Glutathione)
SB  - IM
MH  - Amino Acid Motifs
MH  - Binding Sites
MH  - Cell Cycle
MH  - Cell Line, Tumor
MH  - Glutathione/chemistry/*metabolism
MH  - Glutathione S-Transferase pi/*chemistry/genetics/*metabolism
MH  - Humans
MH  - Kinetics
MH  - Ligands
MH  - Osteosarcoma/enzymology/genetics/*metabolism/physiopathology
MH  - Protein Binding
MH  - TNF Receptor-Associated Factor 2/*chemistry/genetics/*metabolism
PMC - PMC4040697
EDAT- 2014/01/25 06:00
MHDA- 2014/09/03 06:00
PMCR- 2014/01/01
CRDT- 2014/01/25 06:00
PHST- 2013/07/22 00:00 [received]
PHST- 2013/11/08 00:00 [revised]
PHST- 2013/11/13 00:00 [accepted]
PHST- 2014/01/25 06:00 [entrez]
PHST- 2014/01/25 06:00 [pubmed]
PHST- 2014/09/03 06:00 [medline]
PHST- 2014/01/01 00:00 [pmc-release]
AID - cddis2013529 [pii]
AID - 10.1038/cddis.2013.529 [doi]
PST - epublish
SO  - Cell Death Dis. 2014 Jan 23;5(1):e1015. doi: 10.1038/cddis.2013.529.