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Keywords = aminoacyl-tRNA synthetase interacting multi-functional protein 2

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11 pages, 1095 KiB  
Article
Is Methionyl-tRNA Synthetase Applicable as a Diagnostic Marker for Lung Cancer in Bronchial Ultrasound-Guided Brushing Cells?
by Kyungjong Lee, Mijung Oh, Kyo-Sun Lee, Yoon Jin Cha and Yoon Soo Chang
Diagnostics 2021, 11(10), 1830; https://doi.org/10.3390/diagnostics11101830 - 3 Oct 2021
Cited by 3 | Viewed by 2223
Abstract
Background and objective: Methionyl-tRNA synthetase (MARS) and A variant of Aminoacyl-tRNA synthetase interacting multifunctional protein 2 (AIMP2) with an exon 2 deletion (AIMP2-DX2) are known to be overexpressed in lung cancer. However, their role as diagnostic markers in lung cancer has not been [...] Read more.
Background and objective: Methionyl-tRNA synthetase (MARS) and A variant of Aminoacyl-tRNA synthetase interacting multifunctional protein 2 (AIMP2) with an exon 2 deletion (AIMP2-DX2) are known to be overexpressed in lung cancer. However, their role as diagnostic markers in lung cancer has not been well established. Thus, we evaluated their diagnostic performance in brushed cells obtained from nodular lung lesions suspected of lung cancer. Methods: Samples obtained by radial endobronchial ultrasound-guided brushing were processed for cytological examination with Papanicolaou (Pap) staining. Then, double IF staining with MARS and AIMP2-DX2 antibodies was measured in the cytology samples for peripheral lung nodules. The diagnostic performance was compared against biomarkers. Results: MARS IF staining was the only independent staining method used for the prediction of malignant cells. The area under the curve (AUC) of conventional cytology, MARS IF, and MARS IF plus cytology was 0.64, 0.68, and 0.69, respectively. The diagnostic accuracy was increased in MARS IF plus conventional cytology compared with cytology alone (71% vs. 47%). Conclusions: The combination of MARS staining with conventional cytology showed increases in the diagnostic accuracy for diagnosing lung nodules suspected of lung cancer on chest-computed tomography scans. Full article
(This article belongs to the Special Issue Advances in Diagnostic Bronchoscopy)
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11 pages, 1665 KiB  
Article
Anticancer Activity of Pyrimethamine via Ubiquitin Mediated Degradation of AIMP2-DX2
by Dae Gyu Kim, Chul Min Park, Srigouri Huddar, Semi Lim, Sunghoon Kim and Sunkyung Lee
Molecules 2020, 25(12), 2763; https://doi.org/10.3390/molecules25122763 - 15 Jun 2020
Cited by 10 | Viewed by 3743
Abstract
While aminoacyl-tRNA synthetase-interacting multifunctional protein 2 (AIMP2) is a tumor suppressor, its exon 2-depleted splice variant (AIMP2-DX2 or shortly DX2) is highly expressed in human lung cancer, and the ratio of DX2 to AIMP2 increases according to the progression of lung cancer. In [...] Read more.
While aminoacyl-tRNA synthetase-interacting multifunctional protein 2 (AIMP2) is a tumor suppressor, its exon 2-depleted splice variant (AIMP2-DX2 or shortly DX2) is highly expressed in human lung cancer, and the ratio of DX2 to AIMP2 increases according to the progression of lung cancer. In this study, pyrimethamine inhibited the level of DX2 (IC50 = 0.73 µM) in A549 cells expressing nanoluciferase-tagged DX2. In a panel of 5 lung cancer cell lines with various DX2 levels, pyrimethamine most potently suppressed the growth of H460 cells, which express high levels of DX2 (GI50 = 0.01 µM). An immunoblot assay in H460 cells showed that pyrimethamine decreased the DX2 level dose-dependently but did not affect the AIMP2 level. Further experiments confirmed that pyrimethamine resulted in ubiquitination-mediated DX2 degradation. In an in vivo mouse xenograft assay using H460 cells, intraperitoneal administration of pyrimethamine significantly reduced the tumor size and weight, comparable with the effects of taxol, without affecting body weight. Analysis of tumor tissue showed a considerably high concentration of pyrimethamine with a decreased levels of DX2. These results suggest that pyrimethamine, currently used as anti-parasite drug, could be repurposed to treat lung cancer patients expressing high level of DX2. Full article
(This article belongs to the Special Issue Advances in Anticancer Drug Discovery)
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20 pages, 2944 KiB  
Article
An Isoform of the Oncogenic Splice Variant AIMP2-DX2 Detected by a Novel Monoclonal Antibody
by Dae Gyu Kim, Thi Thu Ha Nguyen, Nam Hoon Kwon, Junsik Sung, Semi Lim, Eun-Joo Kang, Jihye Lee, Woo Young Seo, Arum Kim, Yoon Soo Chang, Hyunbo Shim and Sunghoon Kim
Biomolecules 2020, 10(6), 820; https://doi.org/10.3390/biom10060820 - 27 May 2020
Cited by 1 | Viewed by 3748
Abstract
AIMP2-DX2, an exon 2-deleted splice variant of AIMP2 (aminoacyl-tRNA synthetase-interacting multifunctional protein 2), is highly expressed in lung cancer and involved in tumor progression in vivo. Oncogenic function of AIMP2-DX2 and its correlation with poor prognosis of cancer patients have been well established; [...] Read more.
AIMP2-DX2, an exon 2-deleted splice variant of AIMP2 (aminoacyl-tRNA synthetase-interacting multifunctional protein 2), is highly expressed in lung cancer and involved in tumor progression in vivo. Oncogenic function of AIMP2-DX2 and its correlation with poor prognosis of cancer patients have been well established; however, the application of this potentially important biomarker to cancer research and diagnosis has been hampered by a lack of antibodies specific for the splice variant, possibly due to the poor immunogenicity and/or stability of AIMP2-DX2. In this study a monoclonal antibody, H5, that specifically recognizes AIMP2-DX2 and its isoforms was generated via rabbit immunization and phage display techniques, using a short peptide corresponding to the exon 1/3 junction sequence as an antigen. Furthermore, based on mutagenesis, limited cleavage, and mass spectrometry studies, it is also suggested that the endogenous isoform of AIMP2-DX2 recognized by H5 is produced by proteolytic cleavage of 33 amino acids from N-terminus and is capable of inducing cell proliferation similarly to the uncleaved protein. H5 monoclonal antibody is applicable to enzyme-linked immunosorbent assay, immunoblot, immunofluorescence, and immunohistochemistry, and expected to be a valuable tool for detecting AIMP2-DX2 with high sensitivity and specificity for research and diagnostic purposes. Full article
(This article belongs to the Section Cellular Biochemistry)
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14 pages, 1740 KiB  
Article
CRISPR-Cas9 Mediated Telomere Removal Leads to Mitochondrial Stress and Protein Aggregation
by Hyojung Kim, Sangwoo Ham, Minkyung Jo, Gum Hwa Lee, Yun-Song Lee, Joo-Ho Shin and Yunjong Lee
Int. J. Mol. Sci. 2017, 18(10), 2093; https://doi.org/10.3390/ijms18102093 - 3 Oct 2017
Cited by 26 | Viewed by 9795
Abstract
Aging is considered the major risk factor for neurodegenerative diseases including Parkinson’s disease (PD). Telomere shortening is associated with cellular senescence. In this regard, pharmacological or genetic inhibition of telomerase activity has been used to model cellular aging. Here, we employed CRISPR-Cas9 technology [...] Read more.
Aging is considered the major risk factor for neurodegenerative diseases including Parkinson’s disease (PD). Telomere shortening is associated with cellular senescence. In this regard, pharmacological or genetic inhibition of telomerase activity has been used to model cellular aging. Here, we employed CRISPR-Cas9 technology to instantly remove the telomere to induce aging in a neuroblastoma cell line. Expression of both Cas9 and guide RNA targeting telomere repeats ablated the telomere, leading to retardation of cell proliferation. Instant deletion of telomere in SH-SY5Y cells impaired mitochondrial function with diminished mitochondrial respiration and cell viability. Supporting the pathological relevance of cell aging by CRISPR-Cas9 mediated telomere removal, alterations were observed in the levels of PD-associated proteins including PTEN-induced putative kinase 1, peroxisome proliferator-activated receptor γ coactivator 1-α, nuclear respiratory factor 1, parkin, and aminoacyl tRNA synthetase complex interacting multifunctional protein 2. Significantly, α-synuclein expression in the background of telomere removal led to the enhancement of protein aggregation, suggesting positive feed-forward interaction between aging and PD pathogenesis. Collectively, our results demonstrate that CRISPR-Cas9 can be used to efficiently model cellular aging and PD. Full article
(This article belongs to the Special Issue Genome Editing 2018)
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