Product Pathways - Protein Stability
OTUB1 (D8F7) Rabbit mAb #3783
|3783S||100 µl (10 western blots)||---||In Stock||---|
|3783||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||31||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting
Species predicted to react based on 100% sequence homology: Hamster, Bovine, Dog.
Specificity / Sensitivity
OTUB1 (D8F7) Rabbit mAb recognizes endogenous levels of total OTUB1 protein. This antibody does not cross-react with OTUB2 and based upon sequence alignment, is not predicted to cross-react with OTUB1, isoform 2 (ARF-1).
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ile82 of human OTUB1 protein.
Western blot analysis of extracts from various cell lines using OTUB1 (D8F7) Rabbit mAb.
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with Myc/DDK-tagged cDNA expression constructs encoding full-length human OTUB1, isoform1 (hOTUB1-Myc/DDK, +) and full-length human OTUB2 (hOTUB2-Myc/DDK, +), using OTUB1 (D8F7) Rabbit mAb (upper) and DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368 (lower).
Protein ubiquitination and deubiquitination are reversible processes catalyzed by ubiquitinating enzymes (UBEs) and deubiquitinating enzymes (DUBs) (1,2). DUBs are categorized into 5 subfamilies: USP, UCH, OTU, MJD, and JAMM (1,2). The OTU subfamily comprises a group of approximately 100 putative cysteine proteases that are homologous to the ovarian tumor gene product of Drosophila (3). OTUB1 and OTUB2 (OTU domain-containing Ubal-binding proteins) display no significant similarity to any known DUB, but are close homologs and possess an OTU domain that contains conserved cysteine, histidine, and aspartate residues that define the putative catalytic triad of cysteine proteases. Furthermore, sequence analysis of OTUB1 and OTUB2 reveals the presence of putative Ub-interaction motifs (UIMs) and Ub-associated domains (UBAs), which are characteristic of proteins that regulate protein ubiquitination. OTUB1 and OTUB2 also possess a putative nuclear localization signal (NLS) and a consensus LxxLL motif, which mediates the interaction between transcriptional co-activators and nuclear hormone receptors (4).
OTUB1 exists as two isoforms that are generated by alternative splicing; the shorter 31 kDa isoform is ubiquitously expressed, while the longer 35 kDa isoform (ARF-1) has a more restricted expression pattern and is mostly detected in lymphoid organs (5). Biochemical analysis has demonstrated that OTUB1 has a preference for cleaving K48-linked polyubiquitin chains over K63-linked polyubiquitin chains and is capable of cleaving NEDD8, but not SUMO-1, -2, and -3 or ISG15 conjugates (6). OTUB1 isoforms have been implicated in anergy induction in CD4+ T cells by regulating the stability of the E3 ligase GRAIL (5). More recently, OTUB1 was found to bind to and inhibit the E2 activity of UBE2N through a novel mechanism not involving OTUB1 DUB activity, thus compromising the ability of the E3 ligase RNF168 to drive ubiquitination-dependent repair of DNA double-strand lesions (7). OTUB1 also appears to suppress MDM2-dependent ubiquitination of p53 independent of its catalytic activity, primarily by suppressing the activity of the MDM2 cognate E2 UbcH5 (8).
- Nijman, S.M. et al. (2005) Cell 123, 773-86.
- Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613.
- Makarova, K.S. et al. (2000) Trends Biochem Sci 25, 50-2.
- Balakirev, M.Y. et al. (2003) EMBO Rep 4, 517-22.
- Soares, L. et al. (2004) Nat Immunol 5, 45-54.
- Edelmann, M.J. et al. (2009) Biochem J 418, 379-90.
- Nakada, S. et al. (2010) Nature 466, 941-6.
- Sun, X.X. et al. (2012) EMBO J 31, 576-92.
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