Product Pathways - Protein Stability
APC3 (D3I1V) Rabbit mAb #12530
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Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation,
Species predicted to react based on 100% sequence homology: Hamster, Bovine, Dog, Pig, Horse,
Specificity / Sensitivity
APC3 (D3I1V) Rabbit mAb recognizes endogenous levels of total APC3 protein. This antibody does not cross-react with either APC8/CDC23 or APC6/CDC16.
APC3(D3I1V) Rabbit mAb兔单抗识别内源性的总APC3总蛋白。这一抗体与APC8/CDC23或APC6/CDC16均无交叉反应。
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human APC3 protein.
Western blot analysis of extracts from various cell lines using APC3 (D3I1V) Rabbit mAb.Western blot检测多种细胞提取物。
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human APC3 (hAPC3-Myc/DDK; +), using APC3 (D3I1V) Rabbit mAb. Western blot检测mock transfected (-)或转染表达Myc/DDK-tagged全长人APC3 (hAPC3-Myc/DDK; +)。
Immunoprecipitation of APC3 from MCF7 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or APC3 (D3I1V) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using APC3 (D3I1V) Rabbit mAb.Immunoprecipitation检测MCF7细胞提取物，采用Rabbit (DA1E) mAb IgG XP®同型对照#3900 (lane 2) 或APC3 (D3I1V) Rabbit mAb (lane 3)。Lane 1中样品浓度为10%。Western blot检测采用APC3 (D3I1V) Rabbit mAb。
Cell proliferation in all eukaryotic cells depends strictly upon the ubiquitin ligase (E3) activity of the anaphase promoting complex/cyclosome (APC/C), whose main function is to trigger the transition of the cell cycle from metaphase to anaphase. APC/C is a 1.5 MDa protein complex found in the nucleus of interphase cells. This complex diffuses throughout the cytoplasm and associates with parts of the spindle apparatus during mitosis. APC/C performs its various functions by promoting the assembly of polyubiquitin chains on substrate proteins, which targets these proteins for degradation by the 26S proteasome (1,2). In humans, twelve different APC/C subunits have been identified. Like all E3 enzymes, APC/C utilizes ubiquitin residues that have been activated by E1 enzymes and then transferred to E2 enzymes. Indeed APC/C has been shown to transiently interact with UBCH5 and UBCH10 E2 enzymes, in part, via the RING-finger domain-containing subunit, APC11 (3-5). In addition to E2 enzymes, APC/C activity is also strictly dependent upon one of several cofactors that associate with APC/C during specific phases of the cell cycle. The best studied of these are Cdc20 and Cdh1/FZR1, which contain a C-terminal WD40 domain and participate in the recognition of APC/C substrates by interacting with specific recognition elements in these substrates (6), called D-boxes (7) and KEN-boxes (8).
所有真核细胞的细胞增殖严格依赖于泛素连接酶（E3）APC/C的活性，它的主要功能在于激发细胞周期由分裂中期向分裂后期的转变。APC/C是在分裂间期细胞核中发现的一种1.5 MDa大小的蛋白复合物。这一复合物在细胞质中扩散，并在有丝分裂中与部分纺锤体有关。APC/C通过促进底物蛋白聚泛素链的组装而发挥多种功能，主要是通过26S蛋白酶体使这些蛋白降解(1,2)。在人类，已经鉴定出12种不同的APC/C亚基。像所有E3酶一样，APC/C利用已经被E1酶活化的泛素，然后转化为E2酶。确实，APC/C已经被证实短暂地与UBCH5和UBCH10 E2酶相互作用，部分经过包含有环指结构域的亚基APC11 (3-5)。除外E2酶，APC/C的活性严格地依赖于一些辅助因素，与APC/C所处的细胞周期特殊阶段有关。最好的研究是有关Cdc20 和Cdh1/FZR1，包含有C-末端WD40结构域，并通过这些底物(6)中特殊的识别元件D-boxes (7) 和KEN-boxes (8)参与APC/C底物的识别。
Anaphase-promoting complex subunit 3 (APC3) is the human homolog of Saccharomyces cerevisiae CDC27 (9) and, like APC8/CDC23 and APC6/CDC16, is a component of the tetratricopeptide (TPR) subcomplex of the APC/C. It has been demonstrated that the binding of Cdh1/FZR1 to the APC/C depends upon the presence of APC3, implying that APC/C is activated by the association of Cdh1/FZR1 with APC3, which enables APC/C to recognize the D-box of substrates (10,11). APC3 has been shown to be localized to the centrosome at all stages of the mammalian cell cycle, and to the mitotic spindle, suggesting that APC3 plays a critical role for the transition from metaphase to anaphase during mitosis (12). During mitosis, APC3 becomes phosphorylated at numerous sites. This is predicted to change the surface charge distribution significantly such that these modifications could either induce structural changes within the APC/C by altering subunit-subunit interactions or they could change the affinity for molecules that only transiently associate with the APC/C, such as Cdh1/FZR1 (13,14).
APC3是酿酒酵母CDC27的人同源蛋白(9)，同APC8/CDC23和APC6/CDC16一样，是APC/C的TPR亚复合物的组分之一。已经证实，Cdh1/FZR1与APC/C的结合依赖于APC3的存在，表明APC/C由Cdh1/FZR1和APC3共同激活，从而使APC/C识别底物的D-box (10,11)。APC3被证实定位于哺乳动物细胞周期的所有阶段的中心小体以及有丝分裂纺锤体，表明APC3在有丝分裂从分裂中期向分裂后期转变中起重要作用(12)。在有丝分裂中，APC3在多个位点磷酸化。这预示着显著改变细胞表面电荷分布，诸如这样的修饰不仅可以通过调控亚基-亚基相互作用诱导APC/C结构变化，而且可以通过与APC/C短暂地接触改变分子的亲和力，诸如Cdh1/FZR1 (13,14)。
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- Gmachl, M. et al. (2000) Proc Natl Acad Sci U S A 97, 8973-8.
- Leverson, J.D. et al. (2000) Mol Biol Cell 11, 2315-25.
- Kraft, C. et al. (2005) Mol Cell 18, 543-53.
- Glotzer, M. et al. (1991) Nature 349, 132-8.
- Pfleger, C.M. and Kirschner, M.W. (2000) Genes Dev 14, 655-65.
- Tugendreich, S. et al. (1993) Proc Natl Acad Sci U S A 90, 10031-5.
- Vodermaier, H.C. et al. (2003) Curr Biol 13, 1459-68.
- Tugendreich, S. et al. (1995) Cell 81, 261-8.
- Topper, L.M. et al. Cell Cycle 1, 282-92.
- Kraft, C. et al. (2003) EMBO J 22, 6598-609.
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