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Inhibitor kappa B kinase beta binding by inhibitor kappa B kinase gamma

机译:抑制剂kappa B激酶γ与抑制剂kappa B激酶的结合

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Activation of a large multisubunit protein kinase, called the inhibitor kappa B kinase (IKK) complex, is central to the induction of the family of transcription factors nuclear factor kappa B. IKK is comprised of two catalytic subunits, IKK alpha and IKK beta, and a regulatory IKK gamma subunit. It is known that the catalytic IKK beta and regulatory IKK gamma subunits associate through interactions mediated by the N-terminal region of IKK gamma and an 11-mer peptide located near the C-terminus of IKK beta. In this study, we have defined the minimal IKK gamma segment that binds IKK beta and determined the binding affinity of the IKK beta/IKK gamma complex. We identified that the N-terminal segment spanning residues 40-130 of IKK gamma binds the IKK beta C-terminal domain (residues 665-756) with K-d approximate to 25 nM. Several smaller N-terminal IKK gamma deletion mutants within the N-terminal 130 residues, although in some cases retained IKK beta binding activity, showed a tendency to aggregate and formed covalently linked complexes. However, expansion of the C-terminus of these fragments to residue 210 completely changed the solution behavior of the IKK gamma N-terminus without affecting the IKK binding affinity. We also found that the IKK beta C-terminal domain formed a dimer in solution and the basic unit of the IKK beta/IKK gamma complex was a dimer/dimer.
机译:称为抑制剂kappa B激酶(IKK)复合物的大型多亚基蛋白激酶的激活是诱导转录因子核因子kappa B家族的关键。IKK由两个催化亚基IKK alpha和IKK beta组成,以及调节性IKKγ亚基。众所周知,催化性IKK beta和调节性IKKγ亚基通过IKKγ的N末端区域和位于IKK beta的C末端附近的11-mer肽介导的相互作用而缔合。在这项研究中,我们定义了与IKK beta结合的最小IKKγ片段,并确定了IKK beta / IKKγ复合体的结合亲和力。我们确定,跨越IKKγ残基40-130的N末端片段以约25 nM的K-d结合IKK beta C末端域(残基665-756)。 N末端130个残基内的几个较小的N末端IKKγ缺失突变体,尽管在某些情况下保留了IKKβ结合活性,但显示出聚集并形成共价连接的复合物的趋势。但是,这些片段的C末端扩展到残基210完全改变了IKKγN末端的溶液行为,而不影响IKK结合亲和力。我们还发现,IKK beta C末端结构域在溶液中形成二聚体,而IKK beta / IKKγ复合体的基本单位是二聚体/二聚体。

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