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1.
Chinese Journal of Biotechnology ; (12): 53-62, 2024.
Article Dans Chinois | WPRIM | ID: wpr-1008079

Résumé

Abiotic stresses substantially affect the growth and development of plants. Plants have evolved multiple strategies to cope with the environmental stresses, among which transcription factors play an important role in regulating the tolerance to abiotic stresses. Basic leucine zipper transcription factors (bZIP) are one of the largest gene families. The stability and activity of bZIP transcription factors could be regulated by different post-translational modifications (PTMs) in response to various intracellular or extracellular stresses. This paper introduces the structural feature and classification of bZIP transcription factors, followed by summarizing the PTMs of bZIP transcription factors, such as phosphorylation, ubiquitination and small ubiquitin-like modifier (SUMO) modification, in response to abiotic stresses. In addition, future perspectives were prospected, which may facilitate cultivating excellent stress-resistant crop varieties by regulating the PTMs of bZIP transcription factors.


Sujets)
Facteurs de transcription à motif basique et à glissière à leucines/génétique , Maturation post-traductionnelle des protéines , Phosphorylation , Facteurs de transcription/génétique , Stress physiologique/génétique
2.
Chinese Journal of Oncology ; (12): 1362-1368, 2022.
Article Dans Chinois | WPRIM | ID: wpr-969796

Résumé

Objective: To inhibit the stemness maintenance potential of endometrial cancer and increase the sensitivity of endometrial cancer side population cells to chemotherapy drugs by inducing extensive deSUMOylation modification of proteins. Methods: Flow cytometry was used to sort and culture CD133(+) CD44(+) KLE endometrial cancer cell clone spheres. Protein expression level of small ubiquitin-related modifier 1 (SUMO1) and two stemness maintenance genes of tumor side population cells, octamer binding transcription factor-4 (Oct4) and sex determining region Y-box2 (Sox2), were detected by western blotting method. Lentivirus-mediated Sentrin/SUMO-specific proteases 1 (SENP1) gene was stably transfected into KLE side population cells. Western blotting was used to detect the protein expressions of SENP1, SUMO1, Oct4 and Sox2. The clone formation rate was compared between KLE side population cells with or without SENP1 overexpression. Flow cytometry was applied to detect cell cycle changes. 3-(4, 5-Dimethylthiazole-2)-2, 5-diphenyl-tetrazolium bromide (MTT) experiment and flow cytometry apoptosis method were used to detect the chemosensitivity of the side population of endometrial cancer cells to cisplatin. Tumor-bearing mouse models of endometrial cancer were established to detect the effect of SENP1 overexpression on the chemotherapy sensitivity of cisplatin. Results: Compared with CD133(-)CD44(-) KLE cells, CD133(+) CD44(+) KLE side population cells could form clonal spheres and express higher levels of SUMO1, Oct4 and Sox2 proteins (P<0.05). Compared with KLE side population cells that were not transfected with SENP1 gene, the expression level of SENP1 protein in KLE side population cells overexpressing SUMO1、Oct4 and Sox2 were lower. The clonal sphere formation rate was reduced from (25.67±5.44)% to (7.46±1.42)%, and cell cycle shifted from G(0)/G(1) phase to G(2) phase. IC(50) of cisplatin decreased from (55.46±6.14) μg/ml to (11.55±3.12) μg/ml, and cell apoptosis rate increased from (9.76±2.09)% to (16.79±3.44)%. Overexpression of SENP1 could reduce the tumorigenesis rate of KLE side population cells in vivo and increase their chemotherapy sensitivity to cisplatin (P<0.05). Conclusion: Overexpression of SENP1 can induce protein deSUMOylation modification, inhibit the stemness maintenance potential of endometrial cancer side population cells, and enhance their chemotherapy sensitivity, which provides a new reference for gene therapy of endometrial cancer.


Sujets)
Animaux , Femelle , Humains , Souris , Apoptose , Lignée cellulaire tumorale , Cisplatine/pharmacologie , Cysteine endopeptidases/métabolisme , Tumeurs de l'endomètre/génétique , Cellules de population latérale/anatomopathologie , Sumoylation
3.
Chinese Journal of Clinical Pharmacology and Therapeutics ; (12): 285-291, 2021.
Article Dans Chinois | WPRIM | ID: wpr-1015065

Résumé

AIM: To investigate the mechanism of the involvement of SUMO-ylated Androgen receptor (AR) in tamoxifen resistance and the role of SUMO inhibitor ginkgolic acid in resistance. METHODS: Real-Time PCR was used to detect AR mRNA levels in parental cells MCF-7W and drug-resistant cells MCF-7R, AR protein levels and SUMO levels in MCF-7W and MCF-7R cells was performed by western blot, and CB/IP was applied to detect AR interacts with SUMOE3 ligase PIAS1 and HSP27 in MCF-7R cell chromatin, the transcriptional activity of SUMO AR was also evaluated by the fluorescent reporter gene experiment, the CCK-8 method and the trypan blue exclusion method were used to detect cell viability and cell viability respectively. RESULTS: The mRNA and protein expression levels of AR in MCF-7R cells were significantly higher than those in MCF-7W cells (P<0.05), and there was highly SUMOylated AR in MCF-7R cells. Further research found that there had an obvious interaction between AR and SUMO E3 ligase PIAS1 and HSP27, that was, the SUMOylated AR was modified by E3 ligase. Moreover, androgen R1881 could enhance the transcriptional activity of the SUMOylated AR in a concentration-dependent manner. Compared with ginkgo acid alone, 10 μmol/L of ginkgolic acid combined with 10 μmol/L of enzalutamide treated MCF-7R cells for 3 days, the cell number was significantly reduced, and the number of cell death increased significantly (P<0.05). CONCLUSION: The resistance mechanism of tamoxifen may be due to the enhanced AR transcription and activity increased by the hyperactive SUMOylated AR, SUMO inhibitor ginkgolic acid combined with AR antagonist enzalutamide can be a new strategy for the treatment of tamoxifen resistance.

4.
Journal of Veterinary Science ; : e54-2019.
Article Dans Anglais | WPRIM | ID: wpr-758937

Résumé

Brucella is an intracellular pathogen that invades a host and settles in its immune cells; however, the mechanism of its intracellular survival is unclear. Modification of small ubiquitin-related modifier (SUMO) occurs in many cellular activities. E2 conjugating enzyme 9 (Ubc9) is the only reported ubiquitin-conjugating enzyme that links the SUMO molecule with a target protein. Brucella's intracellular survival mechanism has not been studied with respect to SUMO-related proteins and Ubc9. Therefore, to investigate the relationship between Brucella melitensis 16M and SUMO, we constructed plasmids and cells lines suitable for overexpression and knockdown of SUMO1 and Ubc9 genes. Brucella 16M activated SUMO1/Ubc9 expression in a time-dependent manner, and Brucella 16M intracellular survival was inhibited by SUMO1/Ubc9 overexpression and promoted by SUMO1/Ubc9 depletion. In macrophages, Brucella 16M-dependent apoptosis and immune factors were induced by SUMO1/Ubc9 overexpression and restricted by SUMO1/Ubc9 depletion. We noted no effect on the expressions of SUMO1 and Ubc9 in B. melitensis 16M lipopolysaccharide-prestimulated mouse RAW264.7 macrophages. Additionally, intracellular survival of the 16M△VirB2 mutant was lower than that of Brucella 16M (p < 0.05). VirB2 can affect expression levels of Ubc9, thereby increasing intracellular survival of Brucella in macrophages at the late stage of infection. Collectively, our results demonstrate that B. melitensis 16M may use the VirB IV secretion system of Brucella to interact with SUMO-related proteins during infection of host cells, which interferes with SUMO function and promotes pathogen survival in host cells.


Sujets)
Animaux , Souris , Apoptose , Brucella melitensis , Brucella , Facteurs immunologiques , Macrophages , Plasmides
5.
Journal of Jilin University(Medicine Edition) ; (6): 73-76, 2019.
Article Dans Chinois | WPRIM | ID: wpr-742730

Résumé

Objective:To observe the expressions of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells, and to elucidate the their effects in the development of malignant glioma.Methods:The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively.The Cos7cells and the malignant glioma LN443and U343cells were cultured;the Cos7cells were used as normal cell control group, and the LN443and U343cells as malignant glioma cell group.Western blotting method was used to detect the expression levels of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells.Results:In brain tissue, the expression levels of SENP1, SENP2and SENP6proteins in malignant glioma group were higher than those in normal control group (P<0.05) .Compared with normal cell control group, the expression levels of SENP1, SENP2and SENP6proteins in the LN443and U343cells in malignant glioma cell group were significantly increased (P<0.05) .Conclusion:SENP1, SENP2and SENP6proteins highly express in the malignant glioma tissue and cells, and they may play an important role in promoting the occurrence of malignant glioma.

6.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 706-713, 2019.
Article Dans Chinois | WPRIM | ID: wpr-843393

Résumé

Objective: To investigate the regulation of autophagy by SUMO specific peptidase 3 (SENP3, normally called SUMO specific protease 3) in mouse testis. Methods: Immunofluorescence was used to detect the localization of SENP3 in spermatogenic cells and Sertoli cells of testis. Senp3 wild type (Senp3+/+) mice and Senp3 gene knockout heterozygous (Senp3+/-) mice were subjected to starvation treatment to induce autophagy. Testicular tissue proteins were extracted, and the extent of autophagy was detected by Western blotting. The extent of autophagy of Sertoli cells was detected and compared with that of spermatogenic cells in testis by transmission electron microscopy and immunofluorescence. Results: SENP3 mainly localized in the nucleus of Sertoli cells. Compared to Senp3+/+ mice, the extent of starvation-induced autophagy in Sertoli cells of Senp3+/- mice increased. Conclusion: SENP3 can inhibit autophagy in Sertoli cells during nutrient deficiency, which may play a role in controlling the extent of autophagy.

7.
Journal of Jilin University(Medicine Edition) ; (6): 73-76, 2019.
Article Dans Chinois | WPRIM | ID: wpr-841746

Résumé

Objective: To observe the expressions of SENP1, SENP2 and SENP6 proteins in human malignant glioma tissue and cells∗ and to elucidate the their effects in the development of malignant glioma. Methods: The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively. The Cos7 cells and the malignant glioma LN443 and U343 cells were cultured; the Cos7 cells were used as normal cell control group, and the LN443 and U343 cells as malignant glioma cell group. Western blotting method was used to detect the expression levels of SENP1, SENP2 and SENP6 proteins in human malignant glioma tissue and cells. Results: In brain tissue, the expression levels of SENP1,SENP2 and SENP6 proteins in malignant glioma group were higher than those in normal control group ( P<0. 05). Compared with normal cell control group, the expression levels of SENP1,SENP2 and SENP6 proteins in the LN443 and U343 cells in malignant glioma cell group were significantly increased (P<0. 05). Conclusion: SENP1, SENP2 and SENP6 proteins highly express in the malignant glioma tissue and cells,and they may play an important role in promoting the occurrence of malignant glioma.

8.
Motrivivência (Florianópolis) ; 30(56): 120-139, Dez. 2018.
Article Dans Portugais | LILACS | ID: biblio-968719

Résumé

Na colônia japonesa da cidade de Ivoti, no Rio Grande do Sul, destacam-se as práticas corporais de lutas e, dentre estas, o sumô. O presente estudo tem como objetivo investigar como se sucedeu a prática do sumô em Ivoti, desde as primeiras manifestações, na década de 1960, até meados da década de 2010. Para esse fim, foram analisadas fontes impressas, iconográficas e uma fonte oral. A partir do cotejamento e da interpretação das informações foi possível depreender que, através da prática do sumô, de suas representações e adaptações, os nipo-brasileiros de Ivoti buscaram preservar práticas culturais de seus antepassados, vinculando a arte marcial específica, bem como os momentos sociais atrelados a ela, a uma tradição familiar. Neste cenário, a Associação Cultural e Esportiva Nipo-Brasileira de Ivoti (ACENB) constitui-se em local privilegiado para a promoção do sumô, bem como à manutenção e negociação de representações de identidade.


In the Japonese colony of the city Ivoti, Rio Grande do Sul, the corporal practices of wrestling stand out, among them sumo. The present study aims to investigate how the the practice of sumo occurred in Ivoti, since the firsts manifestations, in the 1960s, until the the mid-2010. For this purpose, printed sources, iconographic sources and an oral source were analyzed. From the collating and interpretation of the information it was possible to understand that, through the practice of sumo and its representations and adaptations, the Japanese-Brazilians of Ivoti sought to preserve cultural practices of their ancestors, linking the specific martial art, just as social moments tied to it, to a family tradition. In this scenario, the Associação Cultural e Esportiva Nipo-Brasileira de Ivoti (ACENB) appears as a privileged space to the development of sumo and the maintenance and negotiation of identity representations.


En la colonia japonesa de la ciudad de Ivoti, en Rio Grande do Sul, destacan las prácticas corporales de luchas y, entre ellas, el sumo. El presente estudio tiene como objetivo investigar cómo se sucedió la práctica del sumo en Ivoti, desde las primeras manifestaciones, en la década de 1960, hasta mediados de la década de 2010. Para ese fin, se analizaron fuentes impresas, iconográficas y una fuente oral. A partir del cotejo y de la interpretación de las informaciones fue posible deducir que, a través de la práctica del sumo, de su s representaciones y adaptaciones, los nipo-brasileños de Ivoti buscaron preservar prácticas culturales de de sus antepasados, vinculando el arte marcial específico, así como los momentos sociales articulados con ella, a una tradición familiar. En este escenario, la Associação Cultural e Esportiva Nipo-Brasileira de Ivoti (ACENB) aparece como local privilegiado para la promoción del sumo y el mantenimiento y negociación de representaciones de identidad.


Sujets)
Sports/histoire , Lutte , Émigrants et immigrants , Brésil , Japon
9.
Immune Network ; : e4-2018.
Article Dans Anglais | WPRIM | ID: wpr-740205

Résumé

The initiation of cellular antiviral signaling depends on host pattern-recognition receptors (PRRs)-mediated recognition of viral nucleic acids that are known as classical pathogen-associated molecular patterns (PAMPs). PRRs recruit adaptor proteins and kinases to activate transcription factors and epigenetic modifiers to regulate transcription of hundreds of genes, the products of which collaborate to elicit antiviral responses. In addition, PRRs-triggered signaling induces activation of various inflammasomes which leads to the release of IL-1β and inflammation. Recent studies have demonstrated that PRRs-triggered signaling is critically regulated by ubiquitin and ubiquitin-like molecules. In this review, we first summarize an updated understanding of cellular antiviral signaling and virus-induced activation of inflammasome and then focus on the regulation of key components by ubiquitin and ubiquitin-like molecules.


Sujets)
Épigénomique , Immunité innée , Inflammasomes , Inflammation , Acides nucléiques , Molécules contenant des motifs associés aux pathogènes , Phosphotransferases , Syndrome dysgénésique et respiratoire porcin , Transduction du signal , Facteurs de transcription , Ubiquitine
10.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 348-352, 2018.
Article Dans Chinois | WPRIM | ID: wpr-695668

Résumé

SUMOylation is a dynamically reversible process that needs to be modified by a specific ligase, while its inverse reaction deSUMOylation is catalyzed by a group of SUMO-specific proteases (SENPs). SUMO-modified target protein molecules are closely related to development and disease, especially tumor, metabolism, inflammation and immunity. SENPs play an important role in SUMO protein maturation and deSUMOylation. This review discussed the functions of SENPs in the development and progression of tumors and related mechanisms.

11.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 348-352, 2018.
Article Dans Chinois | WPRIM | ID: wpr-843763

Résumé

SUMOylation is a dynamically reversible process that needs to be modified by a specific ligase, while its inverse reaction deSUMOylation is catalyzed by a group of SUMO-specific proteases (SENPs). SUMO-modified target protein molecules are closely related to development and disease, especially tumor, metabolism, inflammation and immunity. SENPs play an important role in SUMO protein maturation and deSUMOylation. This review discussed the functions of SENPs in the development and progression of tumors and related mechanisms.

12.
Biomedical and Environmental Sciences ; (12): 288-295, 2017.
Article Dans Anglais | WPRIM | ID: wpr-311412

Résumé

<p><b>OBJECTIVE</b>The aim of this study is to determine whether the SUMO4 M55V polymorphism is associated with susceptibility to type 2 diabetes mellitus (T2DM).</p><p><b>METHODS</b>A meta-analysis was performed to detect the potential association of the SUMO4 M55V polymorphism and susceptibility to T2DM under dominant, recessive, co-dominant (homogeneous and heterogeneous), and additive models.</p><p><b>RESULTS</b>A total of eight articles including 10 case-control studies, with a total of 2932 cases and 2679 controls, were included in this meta-analysis. The significant association between the SUMO4 M55V polymorphism and susceptibility to T2DM was observed in the dominant model (GG + GA versus AA: OR = 1.21, 95% CI = 1.05-1.40, P = 0.009), recessive model (GG versus GA + AA: OR = 1.29, 95% CI = 1.07-1.356, P = 0.010), homozygous model (GG versus AA: OR = 1.41, 95% CI = 1.06-1.56, P = 0.001), and additive model (G versus A: OR = 1.18, 95% CI = 1.08-1.29, P = 0.001), and marginally significant in the heterozygous model (GA versus AA: OR = 1.16, 95% CI = 0.98-1.36, P = 0.080). In subgroup analyses, significant associations were observed in the Chinese population under four genetic models excluding the heterozygous model, whereas no statistically significant associations were observed in the Japanese population under each of the five genetic models.</p><p><b>CONCLUSION</b>The meta-analysis demonstrated that the G allele of the SUMO4 M55V polymorphism could be a susceptible risk locus to T2DM, mainly in the Chinese population, while the association in other ethnic population needs to be further validated in studies with relatively large samples.</p>


Sujets)
Humains , Diabète de type 2 , Épidémiologie , Génétique , Prédisposition génétique à une maladie , Épidémiologie , Génétique , Petites protéines modificatrices apparentées à l'ubiquitine , Génétique , Métabolisme
13.
Chinese Journal of Nervous and Mental Diseases ; (12): 170-175, 2017.
Article Dans Chinois | WPRIM | ID: wpr-619873

Résumé

Objective AIM:To study whether the RWD containing sumoylation enhancer (RSUME) enhanced small ubiquitin related modifiers (SUMO) to competitively inhibit Ubiquitin B (UBB)-mediated degradation of hypoxia hypoxia-inducible factor 1α.(HIF-1α) and the invasive pituitary adenomas.Methods The expression of protein and mRNA levels of RSUME,SUMO-1,UBB and HIF-1α were detected by using immunohistochemistry,western blot and qPCR in 38 cases of non-invasive pituitary adenoma,38 cases of invasive pituitary adenomas and 10 cases of normal pituitary capsule.The expression of SUMO-1 was analyzed in different types of pituitary adenomas.Results The protein,and mRNA levels of RSUME,SUMO-HIF-1αwere significantly higher in the invasive pituitary adenomas than in the non-invasive pituitary adenomas and normal pituitary capsule (P<0.01).The protein and mRNA levels of RSUME,SUMO-HIF-1αwas higher in non-invasive pituitary adenomas than in normal pituitary capsule(P<0.01).However,the protein levels of UBB-HIF-1α were significantly lower in the invasive pituitary adenomas than in the non-invasive pituitary adenomas and normal pituitary capsule (P<0.01).The protein of UBB-HIF-1α were lower in non-invasive pituitary adenomas than in normal pituitary capsule (P<0.01).The expression levels of SUMO were not significantly different among different types of pituitary adenomas (P>0.05).Conclusion RSUM may increase pituitary adenomas angiogenesis and promote tumor invasion through enhancement of SUMO of HIF-1α which competitively inhibits ubiquitination of HIF-1 α.

14.
Asian Pacific Journal of Tropical Biomedicine ; (12): 249-252, 2017.
Article Dans Chinois | WPRIM | ID: wpr-511485

Résumé

Objective: To prove probable relations between serum E3 SUMO-protein ligase NSE2 (NSMCE2) concentration, peroxynitrite related to oxidative stress in nephrolithiasis patients. Methods: A total of 60 patients with nephrolithiasis and 50 healthy volunteers were involved in this study. Colorimetric method was used to detect blood urea, creatinine, uric acid, protein, albumin, total antioxidant status, total oxidant status, peroxynitrite, nitric oxide and oxidative stress index. Glutathione, NSMCE2 and superoxide dismutase were measured by ELISA. Results: A significant increase in level of peroxynitrite, total oxidant status, NSMCE2 and oxidative stress index in patients was observed, while total antioxidant status and glutathione were significantly decreased. Conclusions: The study concluded that serum NSMCE2 significantly correlated with peroxynitrite and oxidative stress in patients with nephrolithiasis.

15.
Asian Pacific Journal of Tropical Biomedicine ; (12): 249-252, 2017.
Article Dans Chinois | WPRIM | ID: wpr-950622

Résumé

Objective To prove probable relations between serum E3 SUMO-protein ligase NSE2 (NSMCE2) concentration, peroxynitrite related to oxidative stress in nephrolithiasis patients. Methods A total of 60 patients with nephrolithiasis and 50 healthy volunteers were involved in this study. Colorimetric method was used to detect blood urea, creatinine, uric acid, protein, albumin, total antioxidant status, total oxidant status, peroxynitrite, nitric oxide and oxidative stress index. Glutathione, NSMCE2 and superoxide dismutase were measured by ELISA. Results A significant increase in level of peroxynitrite, total oxidant status, NSMCE2 and oxidative stress index in patients was observed, while total antioxidant status and glutathione were significantly decreased. Conclusions The study concluded that serum NSMCE2 significantly correlated with peroxynitrite and oxidative stress in patients with nephrolithiasis.

16.
Chinese Journal of Biotechnology ; (12): 1395-1407, 2016.
Article Dans Chinois | WPRIM | ID: wpr-243713

Résumé

Basic helix loop helix (bHLH) transcription factor plays an important role in biological processes. Bmsage is a class of bHLH transcription factor highly expressed in the silk gland of Bombyx mori, which is not only involved in the developmental regulation of the silk gland cells at the embryonic period, but also plays a crucial regulatory role during the synthesis of silk protein. However, currently, much of the property and structure of Bmsage is still remained unknown. To study the property, structure and biological role of Bmsage, we constructed several prokaryotic expression vectors of Bmsage fused with NusA, MBP, SUMO, Trx and His tags, respectively, then screened and determined the best soluble expression vector and condition of Bmsage in Escherichia coli combining with the induction temperature and IPTG concentration, and further purified the recombinant Bmsage by Ni-column affinity chromatography according to the established expression condition and characterized its secondary structure using circular dichroism spectra. The results showed that NusA and MBP could significantly enhance the soluble expression of Bmsage in E. coli, but it was difficult to separate Bmsage from these tags. SUMO could not only increase the soluble expression of Bmsage in E. coli to a certain degree, but also be effectively separated from Bmsage. Other tags did not effectively promote the soluble expression of Bmsage in E. coli. Circular dichroism spectra showed that the purified Bmsage had well-defined α-helix structure in solution, indicating that SUMO may promote the correct folding of Bmsage into native-like structure. These work not only establish a foundation for further study of the property, structure and function of Bmsage, but also provide a reference for the expression and purification of other similar proteins.

17.
Electron. j. biotechnol ; 18(4): 302-306, July 2015. ilus
Article Dans Anglais | LILACS | ID: lil-757868

Résumé

Background Overexpression or mutated activation of Fibroblast growth factor receptor 3 (FGFR3) is involved in the pathogenesis of many tumors. More and more studies focus on the potential usage of therapeutic antibodies against FGFR3. Results In this study, a novel single-chain Fv (ScFv) against FGFR3 was prepared and characterized. To achieve the soluble expression, ScFv was fused with Sumo (Small ubiquitin-related modifier) by polymerase chain reaction (PCR), and cloned into pET-20b. The recombinant bacteria were induced by 0.5 mM Isopropyl-ß-d-thiogalactopyranoside (IPTG) for 16 h at 20°C, and the supernatant liquid of Sumo-ScFv was harvested and purified by Ni-NTA chromatography. After being cleaved by the Sumo protease, the recombinant ScFv was released from the fusion protein, and further purified by Ni-NTA chromatography. The purity of ScFv was shown to be higher than 95% and their yield reached 4 mg per liter of bacterial culture. In vitro data showed that ScFv can significantly attenuate FGF9-induced phosphorylation of FGFR3. Conclusion We provide a novel method to produce soluble expression and bioactive functions of ScFv in Escherichia coli.


Sujets)
Récepteur de type 3 des facteurs de croissance fibroblastique/métabolisme , Anticorps à chaîne unique/isolement et purification , Anticorps à chaîne unique/métabolisme , Solubilité , Spectrométrie de masse , Protéines recombinantes , Technique de Western , Escherichia coli
18.
Journal of Xi'an Jiaotong University(Medical Sciences) ; (6): 472-478, 2015.
Article Dans Chinois | WPRIM | ID: wpr-475966

Résumé

Objective To explore the expressions and correlation of RWD containing sumoylation enhancer (RSUME),small ubiquitin-like modifier (SUMO-1),hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor (VEGF)in gliomas of different pathologic grade.Methods We investigated the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA with reverse transcription-polymerase chain reaction (RT-PCR),and investigated the immunohistochemical staining to determine the expressions of SUMO-1,HIF-1α and VEGF in 63 cases of human gliomas of different pathologic grade and 9 cases of normal brain tissues.We studied its correlation with the pathologic grade and the relationship between the expression of RSUME promoter sumoylation and HIF-1α/VEGF pathway in gliomas.Results There were significant differences (P <0.01)in the expressions of RSUME mRNA,HIF-1αmRNA and VEGF mRNA in glioma tissues.With the increasing degree of pathologic grade in tumor specimens,the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA increased markedly (P <0.01 ).There was a positive correlation of the expression levels of RSUME mRNA with HIF-1αmRNA and VEGF mRNA.There were significant differences (P <0.01 )in the expressions of SUMO-1,HIF-1αand VEGF in glioma tissues by immunohistochemical staining.With the ascending of pathologic grade of tumor specimens,the expression levels of SUMO-1,HIF-1α and VEGF increased markedly (P < 0.01 ).There was a positive correlation between the expression level of SUMO-1 and HIF-1α(r =0.857,P <0.01).The Kaplan-Meier analysis and Log-rank test showed significant differences in progress free survival (PFS)between the RSUME high-expression and low-expression groups (χ2 =36.032,P <0.01).Conclusion RSUME may enhance HIF-1α/VEGF pathway through sumoylation in gliomas.It implicates that RSUME is related to angiogenesis in gliomas and can promote tumor invasion and progression,indicating that RSUME can be a novel target in gliomas treatment.

19.
Chinese Journal of Zoonoses ; (12): 203-207, 2015.
Article Dans Chinois | WPRIM | ID: wpr-474518

Résumé

SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a(+ )‐SENPc was transformed into E .coli Rosetta(DE3) ,then the recombinant SENPc protein was expressed by IPTG induction .SDS‐PAGE and Western blot using anti‐His Tag antibody showed that the expression product of SENPc was a fusion protein with a molecular weight of 43 kD .The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SENP protein provide basis for further functional study of SENP .

20.
Military Medical Sciences ; (12): 805-807, 2015.
Article Dans Chinois | WPRIM | ID: wpr-481068

Résumé

Post-transcriptional regulation of prokaryotes is of pivotal importance to the quality control and stability main -tenance of multiple proteins .The ubiquitin and ubiquitin-like proteins-mediated modifications have widely participated in the manipulation of cell cycle control , subcellular localization and cellular metabolisms .Typical ubiquitin like proteins (UBLs) are ubiquitin,SUMO,Nedd8 and FAT10.Recently, the dynamic regulation mechanisms between sumoylation and de-sumoylation have become the cutting edge in the era of ubiquitin-like modifications.In this review, we briefly summa-rized the entire SUMO-mediated modification systems and the intricate balance regulation .We also discussed the counter acting star molecule SENP1 and its relationship with tumor initiation and progression ,immunological regulation and develop-mental homeostasis .

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