Cysteine-179 of I kappa B kinase beta plays a critical role in enzyme activation by promoting phosphorylation of activation loop serines.
- Author:
Mi Sun BYUN
1
;
Jin CHOI
;
Dae Myung JUE
Author Information
1. Department of Biochemistry, College of Medicine, The Catholic University of Korea, Seoul, Korea. dmjue@catholic.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
cysteine;
I?B kinase;
NF-kappa B;
phosphorylation;
protein serine-threonine kinases
- MeSH:
Transfection;
Serine/*metabolism;
Protein Binding;
Phosphorylation;
Mutant Proteins/chemistry;
MAP Kinase Kinase Kinases/metabolism;
I-kappa B Kinase/*chemistry;
Humans;
Hela Cells;
Enzyme Activation/*physiology;
Cysteine/*physiology;
Cells, Cultured;
Catalytic Domain;
Amino Acid Substitution/physiology;
Adenosine Triphosphate/metabolism
- From:Experimental & Molecular Medicine
2006;38(5):546-552
- CountryRepublic of Korea
- Language:English
-
Abstract:
I kappa B kinase beta (IKK beta) subunit of IKK complex is essential for the activation of NF-kappa B in response to various proinflammatory signals. Cys-179 in the activation loop of IKK beta is known to be the target site for IKK inhibitors such as cyclopentenone prostaglandins, arsenite, and antirheumatic gold compounds. Here we show that a mutant IKK beta in which Cys-179 is substituted with alanine had decreased activity when it was expressed in HEK-293 cells, and TNF stimulation did not restore the activity. Phosphorylation of activation loop serines (Ser-177 and Ser-181) which is required for IKK beta activation was reduced in the IKK beta (C179A) mutant. The activity of IKK beta (C179A) was partially recovered when its phosphorylation was enforced by coexpression with mitogen-activated protein kinase kinase kinases (MAPKKK) such as NF-kappa B inducing kinase (NIK) and MAPK/extracellular signal-regulated kinase kinase kinase 1(MEKK1) or when the serine residues were replaced with phospho-mimetic glutamate. The IKK beta (C179A) mutant was normal in dimer formation, while its activity abnormally responded to the change in the concentration of substrate ATP in reaction mixture. Our results suggest that Cys-179 of IKK beta plays a critical role in enzyme activation by promoting phosphorylation of activation-loop serines and interaction with ATP.
