Acetyl-XPO1 (K568) Antibody
Description
Introduction to XPO1 and its Acetylation at K568
XPO1, also known as Chromosome Region Maintenance 1 protein homolog (CRM1) or Exportin-1, is a crucial protein that mediates leucine-rich nuclear export signal (NES)-dependent protein transport . This cell-cycle-regulated protein plays essential roles in controlling several cellular processes by regulating the localization of important molecules such as cyclin B, MPAK, and MAPKAP kinase 2 . Additionally, XPO1 regulates transcription factors including NFAT and AP-1, and specifically inhibits the nuclear export of Rev and U snRNAs .
The XPO1 gene is conserved across species, with identified homologs in humans, mice, and rats. In the human genome, it is identified by the Entrez Gene ID 7514, while mouse and rat homologs correspond to IDs 103573 and 85252, respectively . The protein products are registered in the Swiss-Prot database under identifiers O14980 (human), Q6P5F9 (mouse), and Q80U96 (rat) .
Acetylation at lysine 568 (K568) represents a specific post-translational modification of XPO1 that may significantly influence its function in nuclear-cytoplasmic transport. This modification site is located within a functionally important region of the protein, potentially affecting its binding capabilities and regulatory properties.
The Acetyl-XPO1 (K568) Antibody is engineered to detect endogenous levels of XPO1 protein specifically when it's acetylated at lysine 568 . Some antibody variants are designed to detect the protein only when non-acetylated at K568, highlighting the importance of verifying the specific detection profile when selecting an antibody for research purposes .
Based on sequence homology, these antibodies typically demonstrate cross-reactivity with human, mouse, and rat XPO1 proteins . This cross-species reactivity makes them valuable tools for comparative studies across different model systems.
Western Blotting
Western blotting represents one of the primary applications for the Acetyl-XPO1 (K568) Antibody . For this technique, manufacturers typically recommend dilutions ranging from 1:500 to 1:2000, depending on the specific antibody preparation and experimental conditions .
In Western blot analysis, the antibody detects a band of approximately 123-125 kDa, corresponding to the full-length acetylated XPO1 protein . Experimental validation has demonstrated successful detection in various sample types, including UV-treated HepG2 cell lysates and mouse brain whole cell lysates . The slight difference between the predicted size (123 kDa) and observed size (125 kDa) likely reflects post-translational modifications that affect protein migration in SDS-PAGE.
Additional Applications
Beyond Western blotting, the Acetyl-XPO1 (K568) Antibody has demonstrated utility in enzyme-linked immunosorbent assays (ELISA) with recommended dilutions of approximately 1:10000 . The high dilution factor for ELISA applications suggests strong binding affinity and specificity.
While not explicitly detailed in the available search results, antibodies of this type are often also applicable for immunohistochemistry, immunocytochemistry, and immunoprecipitation studies, though optimization may be required for these applications.
Table 2: Recommended Applications and Dilutions
| Application | Typical Dilution Range | Detection Notes |
|---|---|---|
| Western Blotting | 1:500 - 1:2000 | Detects band at ~123-125 kDa |
| ELISA | 1:10000 | High sensitivity |
| Immunohistochemistry | Protocol-dependent | May require optimization |
| Immunoprecipitation | Protocol-dependent | May require optimization |
XPO1 in Disease Research
XPO1 has emerged as a significant target in cancer research, with studies demonstrating its involvement in various malignancies. Selective Inhibitor of Nuclear Export (SINE) compounds, such as KPT-330 (selinexor) and KPT-8602, have been developed to target XPO1 by covalently binding to Cys528 in the NES-binding groove, preventing cargo binding and export .
The Acetyl-XPO1 (K568) Antibody provides researchers with a powerful tool to investigate how acetylation at K568 might influence XPO1 function and its response to inhibitors. This is particularly relevant as research has shown that XPO1 inhibitors can induce proteasome-mediated degradation of XPO1, a phenomenon that might be influenced by its acetylation status .
Cellular Response Studies
Studies have demonstrated that XPO1 inhibition can have significant effects on cellular processes in cancer cells. For instance, treatment with SINE compounds leads to nuclear accumulation of tumor suppressor proteins like p53 and NPM1, which are normally exported from the nucleus by XPO1 . The Acetyl-XPO1 (K568) Antibody enables researchers to investigate whether acetylation at K568 affects these critical cargo interactions.
Research utilizing UV-treated cells has shown modulation of XPO1 acetylation patterns, suggesting potential connections between cellular stress responses and XPO1 post-translational modifications . This opens avenues for investigating how various cellular stimuli might regulate XPO1 function through acetylation.
Experimental Controls
When working with the Acetyl-XPO1 (K568) Antibody, appropriate controls should be included to validate results. Positive controls might include UV-treated HepG2 cells, which have been shown to exhibit detectable levels of acetylated XPO1 . Negative controls might involve samples treated with deacetylase inhibitors or competing peptides to confirm specificity.
Future Research Directions
The Acetyl-XPO1 (K568) Antibody opens several promising avenues for future research:
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Investigating the relationship between XPO1 acetylation status and its sensitivity to SINE compounds and other inhibitors
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Examining how acetylation at K568 affects XPO1's interaction with different cargo proteins under normal and disease conditions
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Exploring potential crosstalk between acetylation and other post-translational modifications of XPO1
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Developing therapeutic strategies that specifically target acetylated forms of XPO1 in diseases where this modification plays a role
Understanding the complex regulatory mechanisms governing XPO1 acetylation may ultimately contribute to more refined therapeutic approaches targeting the nuclear export machinery in cancer and other diseases.
Product Specs
Target Background
- We describe three in vitro reconstituted disassembly intermediates, which show binding of a Crm1 export complex via two FG-repeat patches, cargo-release by RanBP2's Ran-binding domains and retention of free Crm1 at RanBP2 after Ran-GTP hydrolysis. PMID: 27160050
- Nuclear entrapment of p33ING1b by inhibition of exportin-1 triggers apoptosis in head and neck squamous cell cancer cells. PMID: 29729696
- CDK4 and XPO1 are not altered in a rare undifferentiated sarcoma, making them therapeutic targets PMID: 27329820
- The subcellular distributions of IkappaB and NFkappaB are indicative of carcinogenesis. Inhibition of XPO1 results in intranuclear retention of IkappaB, which inhibits NFkappaB and thereby provides a novel mechanism for drug therapy in sarcoma. This effect can be further enhanced in relatively selinexor-resistant sarcoma cell lines by pretreatment with the proteasome inhibitor carfilzomib. PMID: 28314790
- This work advocates for assessing 2p+ and XPO1 mutations before choosing a chronic lymphocytic leukemia therapy. PMID: 28344316
- Importin-beta and CRM1 control a RANBP2 spatiotemporal switch essential for mitotic kinetochore function. PMID: 28600321
- We provide evidence for a regulatory role of CRM1 (chromosome-region-maintenance-1; also known as XPO1, exportin-1) in juxta-nuclear microtubule-dependent adenovirus transport. Leptomycin B (LMB) abolishes nuclear targeting of adenovirus. It binds to CRM1, precludes CRM1-cargo binding and blocks signal-dependent nuclear export. PMID: 28515232
- In leukemia cell lines an XPO1 heterozygous mutation confers similar resistance against selinexor as homozygous substitution, demonstrating that SINE resistance can be obtained by a single and dominant mutation of the cysteine528 residue in XPO1 PMID: 27634897
- XPO1 inhibitor combination therapy with bortezomib or carfilzomib induces nuclear localization of IkappaBalpha and overcomes acquired proteasome inhibitor resistance in human multiple myeloma. PMID: 27806331
- KPT-8602 is highly specific for XPO1 inhibition and demonstrates potent anti-leukemic activity supporting clinical application of the second-generation SINE compound for the treatment of Acute Lymphoblastic Leukemia PMID: 27780859
- Taken together, these results provide evidence that XPO1 inhibition represents a new therapeutic strategy for overcoming platinum resistance in women with ovarian cancer PMID: 27649553
- Selinexor, a selective inhibitor of XPO1, is currently being tested as single agent in clinical trials in acute myeloid leukemia. PMID: 27358488
- Nuclear export receptor CRM1 recognizes diverse conformations in nuclear export signals. PMID: 28282025
- These results suggest a differential interaction between human Crm1 and mouse Crm1 and many lentiviral Rev proteins, which may partially explain the HIV replicative defect in mice. PMID: 29028476
- Combined targeting of XPO1 and ERalpha in several tamoxifen-resistant cell lines and tumor xenografts with the XPO1 inhibitor, Selinexor, and tamoxifen restored tamoxifen sensitivity and prevented recurrence in vivo. PMID: 27533791
- Results suggest that the cancer-inhibitory activity of sodium butyrate and its derivatives on liver carcinogenesis may be attributed to retention of p53 and CRM1 proteins in the nucleus, an event that may trigger activation of p53-mediated apoptotic cell death in neoplastic cells. PMID: 27013579
- REVIEW: the role of XPO1 in B cell hematological malignancies PMID: 28196522
- Selinexor, an inhibitor of XPO1, induces cell cycle arrest independent of alterations in the KIT signaling pathway. PMID: 26918731
- Authors investigated the clinical significance of XPO1 mutations in patients with CLL. PMID: 27468087
- Here, the authors identify cellular nuclear transport factor 2 (NTF2)-like export protein 1 (NXT1) as a novel binding partner of nucleoprotein (NP) that stimulates NP-mediated nuclear export via the CRM1-dependent pathway. PMID: 27483302
- Data show that the cytoplasmic localization of promyelocytic leukaemia (PML) is mediated by its nuclear export in a chromosomal maintenance 1 (CRM1)-dependent manner. PMID: 26549027
- CRM1 and CDK5 co-expression was an independent prognostic factors for gastric cancer (GC). Combined CRM1 and CDK5 expression could provide a prognostic model for overall survival of GC. PMID: 28373767
- Anti-tumor activity of selective exportin 1 inhibitors is enhanced in non-Hodgkin lymphoma through combination with mTOR inhibitor and dexamethasone. PMID: 27693556
- The functional consequences of a recurrent cancer-related mutation, which targets a residue near CRM1 NES-binding cleft, was investigated. PMID: 27312238
- We characterized the biologic significance of CRM1 in the context of Ewing sarcoma and determined the therapeutic merit of CRM1 inhibition for this malignancy. PMID: 26956669
- XPO1 E571K mutation represents a genetic hallmark of the PMBL subtype and serves as a new relevant PMBL biomarker. SINE compounds appear active for both mutated and wild-type protein PMID: 27312795
- CRM1 mediates nuclear export of influenza A nucleoprotein. PMID: 28399435
- The detection of the XPO1 E571K mutation in biopsy and plasma cell-free DNA by digital PCR may be used as a novel biomarker in classical Hodgkin lymphoma for both diagnosis and minimal residual disease, and pinpoints a crucial role of XPO1 in classical Hodgkin lymphoma pathogenesis. PMID: 27479820
- The study demonstrated the association of NUP98-IQCG with CRM1, and found that NUP98-IQCG expression inhibits the CRM1-mediated nuclear export of p65 and enhances the transcriptional activity of nuclear factor-kappaB. Moreover, IQCG could be entrapped in the nucleus by NUP98-IQCG, and the fusion protein interacts with calmodulin via the IQ motif in a calcium-independent manner. PMID: 27864780
- XPO1 inhibition has downstream effects on the 3D nuclear organization of the genome. PMID: 26991404
- Findings suggest that mitotic abnormalities can be prevented by the modulation of CRM1 and survivin. We demonstrated the ability of compound 'IV08.009' to efficiently protect cultured keratinocytes from mitotic abnormalities PMID: 26859314
- The interplay between CRM-1 and p27Kip1 may provide potentially potent biomarkers and functional targets for the development of future cholangiocarcinoma treatments. PMID: 27279267
- Data show that functional Exportin 1 (XPO1/CRM1) inhibition correlates to XPO1 occupancy by selinexor in U2OS cells. PMID: 26654943
- The results indicate that highly selective targeting of Nup98-fusion proteins to Hox cluster regions via prebound Crm1 induces the formation of higher order chromatin structures that causes aberrant Hox gene regulation. PMID: 26740045
- Together, our study identifies CRM1 as a valid target in ovarian cancer and provides a basis for the development of S109 in ovarian cancer. PMID: 26055813
- This regulation was conserved in HIV-2 and was dependent on the CRM1-dependent nuclear export pathway suggesting a role of the RNA helicase in interconnecting nuclear export with ribosome recruitment of the viral unspliced mRNA PMID: 27012366
- HIV-1 depends on host-cell-encoded factors to complete its life cycle; data suggest nucleus-located NAF1 (HIV Nef-associated factor 1) promotes nuclear export of un-spliced HIV-1 gag mRNA; association between NAF1 and CRM1 is required for this function. PMID: 26733199
- The binding of nuclear export signals to CRM1 in both orientations results in a large expansion in nuclear export signal consensus patterns and therefore a corresponding expansion of potential nuclear export signals in the proteome. PMID: 26349033
- Findings indicate that exportin 1 protein (CRM1) is a valid target for the treatment of colorectal cancer. PMID: 25996664
- Our results elucidate that RanGAP1 is actively transported between the nuclear and cytoplasmic compartments, and that the cytoplasmic and NPC localization of RanGAP1 is dependent on CRM1-mediated nuclear export. PMID: 26506250
- Ribosomal biogenesis appears to be a key component through which XPO1 contributes to tumor cell survival. PMID: 26340096
- review of physiological function of chromosome region maintenance 1 protein. PMID: 26048327
- Our study suggests SINE-mediated XPO1/CRM1 inhibition as a novel therapeutic option for DMPM. PMID: 25948791
- These data suggest that CRM1 plays an important role in lung carcinogenesis. PMID: 25629636
- The export receptor Crm1 forms a dimer to promote nuclear export of HIV RNA. PMID: 25486595
- This study identifies a cellular protein named RBM14 that is associated with XPO1 (CRM1), a nuclear protein that binds to the HIV-1 Rev protein and mediates nuclear export of incompletely spliced HIV-1 viral RNAs PMID: 25589658
- The new CRM1 inhibitors, therefore, hold strong potential and warrant further clinical investigations for PDAC. PMID: 24899509
- CRM1 has a role in regulating HOXA gene transcription in CALM-AF10 leukemias PMID: 25027513
- Studied CRM1 expression in esophageal squamous cell carcinoma; statistical analysis demonstrated that patients with high CRM1 levels indicated shorter survival period. We further found that silencing CRM1 caused apoptosis in ESCC cell lines. PMID: 25148895
- CRM1 as a new therapeutic target for non-Hodgkin lymphoma. PMID: 25466285
