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FSCB phosphorylation in mouse spermatozoa capacitation
( Shun Li Liu ),( Bing Ni ),( Xiang Wei Wang ),( Wen Qian Huo ),( Jun Zhang ),( Zhi Qiang Tian ),( Ze Min Huang ),( Yi Tian ),( Jun Tang ),( Yan Hua Zheng ),( Feng Shuo Jin ),( Yan Feng Li ) 생화학분자생물학회(구 한국생화학분자생물학회) 2011 BMB Reports Vol.44 No.8
It is generally accepted that spermatozoa capacitation is associated with protein kinase A-mediated tyrosine phosphorylation. In our previous study, we identified the fibrous sheath CABYR binding protein (FSCB), which was phosphorylated by PKA. However, the phosphorylation status of FSCB protein during spermatozoa capacitation should be further investigated. To this aim, in this study, we found that phosphorylation of this 270-kDa protein occurred as early as 1 min after mouse spermatozoa capacitation, which increased over time and remained stable after 60 min. Immunoprecipitation assays demonstrated that the tyrosine and Ser/Thr phosphorylation of FSCB occurred during spermatozoa capacitation. The extent of phosphorylation and was closely associated with the PKA activity and spermatozoa motility characteristics. FSCB phosphorylation could be induced by PKA agonist DB-cAMP, but was blocked by PKA antagonist H-89.Therefore, FSCB contributes to spermatozoa capacitation in a tyrosine-phosphorylated format, which may help in further elucidating the molecular mechanism of spermatozoa capacitation. [BMB reports 2011; 44(8): 541-546]
( Ze Min Huang ),( Zheng Cai Jia ),( Jun Tang ),( Yi Zhang ),( Yi Tian ),( Dong Jing Ni ),( Fang Wang ),( Yu Zhang Wu ),( Bing Ni ) 생화학분자생물학회(구 한국생화학분자생물학회) 2012 BMB Reports Vol.45 No.7
Almost all melanoma cells express at least one member of the MAGE-A antigen family, making the cytotoxic T cells (CTLs) epitopes with cross-immunizing potential in this family attractive candidates for the broad spectrum of anti-melanoma immunotherapy. In this study, four highly homologous peptides (P264: FLWGPRALA, P264I9: FLWGPRALI, P264V9: FLWGPRALV, and P264H8: FLWGPRAHA) from the MAGE-A antigens were selected by homologous alignment. All four peptides showed high binding affinity and stability to HLA-A*02:01 molecules, and could prime CTL immune responses in human PBMCs and in HLA-A*02:01/Kb transgenic mice. CTLs elicited by the four epitope peptides could cross-lyse tumor cells expressing the mutual target antigens, except MAGE-A11 which was not tested. However, CTLs induced by P264V9 and P264I9 showed the strongest target cell lysis capabilities, suggesting both peptides may represent the common CTL epitopes shared by the eight MAGE-A antigens, which could induce more potent and broad-spectrum antitumor responses in immunotherapy. [BMB Reports 2012; 45(7): 408-413]
( Ze Min Huang ),( Jun Wu ),( Zheng Cai Jia ),( Yi Tian ),( Jun Tang ),( Yan Tang ),( Ying Wang ),( Yu Zhang Wu ),( Bing Ni ) 생화학분자생물학회 (구 한국생화학분자생물학회) 2012 BMB Reports Vol.45 No.6
The retinoid-related orphan nuclear receptor gamma (RORγ) plays critical roles in regulation of development, immunity and metabolism. As transcription factor usually forms a protein complex to function, thus capturing and dissecting of the RORγ protein complex will be helpful for exploring the mechanisms underlying those functions. After construction of the recombinant tandem affinity purification (TAP) plasmid, pMSCVpuro RORγ-CTAP(SG), the nuclear localization of RORγ-CTAP(SG) fusion protein was verified. Following isolation of RORγ protein complex by TAP strategy, seven candidate interacting proteins were identified. Finally, the heat shock protein 90 (HSP90) and receptor-interacting protein 140 (RIP140) were confirmed to interplay with RORγ by co-immunoprecipitation. Interference of HSP90 or/and RIP140 genes resulted in dramatically decreased expression of CYP2C8 gene, the RORγ target gene. Data from this study demonstrate that HSP90 and RIP140 proteins interact with RORγ protein in a complex format and function as co-activators in the RORγ-mediated regulatory processes of HepG2 cells. [BMB Reports 2012; 45(6): 331-336]
Ze-Qin Dai,Hang Su,Min Luo,Yu Ou,Xiao-Zhong Fu,Yong-Xi Dong,Yu-Feng Cha,Shun Zhang,Yong Huang,Yong-Lin Wang 대한화학회 2015 Bulletin of the Korean Chemical Society Vol.36 No.8
A series of 4′-N-substituted (aminomethyl)benzoate derivatives of scutellarein were designed and synthesized. Evaluation of their physiochemical properties showed that the designed target compounds 5a–e exhibit higher chemical stability and aqueous solubility than scutellarin and scutellarein. The permeability (Papp AP to BL ) of 5c–e in Caco-2 cells were 2.8, 8.1, and 12.6 times higher than that of scutellarin and 1.3, 4.1, and 6.0 times higher than that of scutellarein; especially, 5e had the highest P app AP to BL value (7.19 ± 0.31 × 10−6 cm/s) and the lowest ER (P app BL to AP /P app AP to BL ) value of 0.17. In vitro antioxidative evaluation results revealed that 5e could protect against H2O2 -induced PC12 cells’ oxidative damage by attenuating mitochondrial membrane potential loss and decreasing H2O2 -induced reactive oxygen species (ROS) production.
Zhi, Ai-Min,Feng, Ding-Yuan,Zhou, Xiang-Yan,Zou, Shi-Geng,Huang, Zhi-Yi,Zuo, Jian-Jun,Ye, Hui,Zhang, Chang-Ming,Dong, Ze-Min,Liu, Zhun Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.8
Cationic amino acid transporter $b^{0,+}AT$ (HGMW-approved gene symbol SLC7A9, solute carrier family 7, member 9) plays a crucial role in amino acid nutrition. In the present study, we describe the cloning and sequencing of porcine $b^{0,+}AT$. Based on the sequence of porcine $b^{0,+}AT$ deposited in the NCBI (National Center for Biotechnological Information), we identified a putative porcine homologue. Using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine $b^{0,+}AT$ was isolated. The porcine $b^{0,+}AT$ cDNA was 1,680 bp long, encoding a 487 amino acid trans-membrane protein. The predicted amino acid sequence was found to have 88.9% and 87.1% identity with human and mouse $b^{0,+}AT$, respectively. Real-time RT-PCR indicated porcine $b^{0,+}AT$ transcripts expressed in heart, kidney, muscle and small intestine. The small intestine had the highest $b^{0,+}AT$ mRNA abundance while the muscle had the lowest (p<0.05). Along the longitudinal axis, the ileum had the highest $b^{0,+}AT$ mRNA abundance while the colon had the lowest (p<0.05). The $b^{0,+}AT$ mRNA level was highest on day 7 and 90 in the duodenum (p<0.05). It increased from day 1 to day 26 in the jejunum (p>0.05) and had the highest abundance on day 60 (p<0.05). There was, however, no difference between day 1, 7, 26, 30, 90 and 150 (p>0.05). The strongest $b^{0,+}AT$ expression appeared on day 7 in the ileum before weaning, and then decreased till day 30 but rose gradually again from day 60 to 150 (p<0.05).