Structural model of ubiquitin transfer onto an artificial RING finger as an E3 ligase
نویسنده
چکیده
The artificial WSTF PHD_EL5 RING finger was designed via "α-helical region substitution", and its structural model for the attachment of activated ubiquitin has been demonstrated. Chemical modifications of Cys residues, the circular dichroism spectra, and substrate-independent ubiquitination assays illustrated that the WSTF PHD_EL5 RING finger has E3 activity, and it is ubiquitinated via Lys14. Homology modeling calculations revealed that the WSTF PHD_EL5 RING finger possesses a classical RING fold for specific E2-E3 binding. The docking poses of the WSTF PHD_EL5 RING finger with the UbcH5b-ubiquitin conjugate provided insight into its functional E2 interaction and development of ubiquitination at the atomic level. The structural model of the artificial WSTF PHD_EL5 RING finger proposed by the present work is useful and may help to extend the strategy of α-helical region substitution.
منابع مشابه
Conserved function of RNF4 family proteins in eukaryotes: targeting a ubiquitin ligase to SUMOylated proteins.
The function of small ubiquitin-like modifier (SUMO)-binding proteins is key to understanding how SUMOylation regulates cellular processes. We identified two related Schizosaccharomyces pombe proteins, Rfp1 and Rfp2, each having an N-terminal SUMO-interacting motif (SIM) and a C-terminal RING-finger domain. Genetic analysis shows that Rfp1 and Rfp2 have redundant functions; together, they are e...
متن کاملRole of Oxidative Stress, ER Stress and Ubiquitin Proteasome System in Neurodegeneration
NDD: Neurodegenerative Disorders; ER: Endoplasmic Reticulum; UPS: Ubiquitin Proteasome System; ROS: Reactive Oxygen Species; AD: Alzheimer’s Disease; PD: Parkinson’s Disease; MS: Multiple Sclerosis; HD: Huntington’s Disease; ALS: Amyotrophic Lateral Sclerosis; PS: Presenilin; APP: Amyloid beta (A4) Precursor Protein; MARK1: Microtubule Affinity-Regulating Kinase 1; SOD-1: Superoxide Dismutase 1...
متن کاملStructure of the HHARI Catalytic Domain Shows Glimpses of a HECT E3 Ligase
The ubiquitin-signaling pathway utilizes E1 activating, E2 conjugating, and E3 ligase enzymes to sequentially transfer the small modifier protein ubiquitin to a substrate protein. During the last step of this cascade different types of E3 ligases either act as scaffolds to recruit an E2 enzyme and substrate (RING), or form an ubiquitin-thioester intermediate prior to transferring ubiquitin to a...
متن کاملHerpes simplex virus type 1 immediate-early protein ICP0 and is isolated RING finger domain act as ubiquitin E3 ligases in vitro.
Proteasome-dependent degradation of ubiquitinated proteins plays a key role in many important cellular processes. Ubiquitination requires the E1 ubiquitin activating enzyme, an E2 ubiquitin conjugating enzyme, and frequently a substrate-specific ubiquitin protein ligase (E3). One class of E3 ubiquitin ligases has been shown to contain a common zinc-binding RING finger motif. We have previously ...
متن کاملEngineering a ubiquitin ligase reveals conformational flexibility required for ubiquitin transfer.
Protein ubiquitination regulates numerous cellular functions in eukaryotes. The prevailing view about the role of RING or U-box ubiquitin ligases (E3) is to provide precise positioning between the attached substrate and the ubiquitin-conjugating enzyme (E2). However, the mechanism of ubiquitin transfer remains obscure. Using the carboxyl terminus of Hsc70-interacting protein as a model E3, we s...
متن کامل