Role of cysteine residues of p65/NF-kappaB on the inhibition by the sesquiterpene lactone parthenolide and N-ethyl maleimide, and on its transactivating potential
Standard
Role of cysteine residues of p65/NF-kappaB on the inhibition by the sesquiterpene lactone parthenolide and N-ethyl maleimide, and on its transactivating potential. / García-Piñeres, A J; Lindenmeyer, M T; Merfort, I.
in: LIFE SCI, Jahrgang 75, Nr. 7, 02.07.2004, S. 841-56.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Role of cysteine residues of p65/NF-kappaB on the inhibition by the sesquiterpene lactone parthenolide and N-ethyl maleimide, and on its transactivating potential
AU - García-Piñeres, A J
AU - Lindenmeyer, M T
AU - Merfort, I
N1 - Copyright 2004 Elsevier Inc.
PY - 2004/7/2
Y1 - 2004/7/2
N2 - Sesquiterpene lactones (SLs) are potent anti-inflammatory substances. It was previously shown that the anti-inflammatory effect could be partly explained by the inhibition of the transcription factor NF-kappaB. Whether they inhibit the DNA binding of NF-kappaB, the activation of the IkappaB-kinase, or both is still a matter of debate. The data supporting these hypotheses were obtained using different cell systems. In this contribution we analyzed the mechanism of the sesquiterpene lactone-mediated inhibition using different cell systems, and showed that in all the cell lines analyzed, SLs inhibited both NF-kappaB binding and the IkappaB-kinase, but that the former played a more preponderant role in the inhibition. These results again confirm the importance of cysteine 38 in the inhibition and regulation of NF-kappaB's function. Moreover, we compared the selectivity of the SL parthenolide with that of N-ethyl maleimide (NEM). We showed that NEM directly alkylated p65 as well as p50 of NF-kappaB, whereas SLs possess a selectivity towards p65. Finally, we studied the transactivating properties of various p65 mutants, to analyze the effect of exchanged cysteine residues in the DNA binding domain of NF-kappaB/p65 on its function and demonstrated that the transactivating potential of the mutants did not correlate with their DNA binding strenght.
AB - Sesquiterpene lactones (SLs) are potent anti-inflammatory substances. It was previously shown that the anti-inflammatory effect could be partly explained by the inhibition of the transcription factor NF-kappaB. Whether they inhibit the DNA binding of NF-kappaB, the activation of the IkappaB-kinase, or both is still a matter of debate. The data supporting these hypotheses were obtained using different cell systems. In this contribution we analyzed the mechanism of the sesquiterpene lactone-mediated inhibition using different cell systems, and showed that in all the cell lines analyzed, SLs inhibited both NF-kappaB binding and the IkappaB-kinase, but that the former played a more preponderant role in the inhibition. These results again confirm the importance of cysteine 38 in the inhibition and regulation of NF-kappaB's function. Moreover, we compared the selectivity of the SL parthenolide with that of N-ethyl maleimide (NEM). We showed that NEM directly alkylated p65 as well as p50 of NF-kappaB, whereas SLs possess a selectivity towards p65. Finally, we studied the transactivating properties of various p65 mutants, to analyze the effect of exchanged cysteine residues in the DNA binding domain of NF-kappaB/p65 on its function and demonstrated that the transactivating potential of the mutants did not correlate with their DNA binding strenght.
KW - Animals
KW - Anti-Inflammatory Agents, Non-Steroidal
KW - Calcium-Binding Proteins
KW - Cysteine
KW - DNA-Binding Proteins
KW - Ethylmaleimide
KW - HeLa Cells
KW - Humans
KW - I-kappa B Kinase
KW - Jurkat Cells
KW - Macrophages
KW - Membrane Glycoproteins
KW - Mice
KW - Mutation
KW - NF-kappa B
KW - Nerve Tissue Proteins
KW - Protein-Serine-Threonine Kinases
KW - Sesquiterpenes
KW - Synaptotagmin I
KW - Synaptotagmins
KW - Transcription Factor RelA
KW - Transcriptional Activation
KW - Transfection
KW - Comparative Study
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1016/j.lfs.2004.01.024
DO - 10.1016/j.lfs.2004.01.024
M3 - SCORING: Journal article
C2 - 15183076
VL - 75
SP - 841
EP - 856
JO - LIFE SCI
JF - LIFE SCI
SN - 0024-3205
IS - 7
ER -