Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica最新文献_第9页

[Mapping of FHL2 transcription activation domain]. [FHL2转录激活域的定位]。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Jing-Hua Yan, Qi-Nong Ye, Yan Fang, Jian-Hua Zhu, Cui-Fen Huang

{"title":"[Mapping of FHL2 transcription activation domain].","authors":"Jing-Hua Yan, Qi-Nong Ye, Yan Fang, Jian-Hua Zhu, Cui-Fen Huang","doi":"","DOIUrl":"","url":null,"abstract":"FHL2, a member of LIM-only protein family, plays an important role in transcription regulation, apoptosis, cancer development and progression. In this study, a mammalian transcription activation system was constructed by using DNA binding domain(DBD) of GAL4 and luciferase reporter gene with DBD binding sequence, and used for mapping of FHL2 transcription activation domain. First, the coding sequence of GAL4-DBD was inserted into expression vector pcDNA3, generating the pDBD recombinant plasmid, then the wild-type FHL2 and its mutants were fused in-frame with GAL4-DBD, resulting in expression vectors for wild-type FHL2 and its mutants. All of the recombinant plasmids were transfected into 293T cells. Western blot assay showed that all of the fusion proteins were expressed. The analysis of FHL2 transcription activation properties by using the GAL4-luciferase reporter gene indicated that wild-type FHL2 had activation activity in both 293T and MCF-7 cells. The deletion of the half LIM domain at the N-terminus severely impaired the capacity of FHL2 to stimulate transcription. The mutant lacking the LIM domain at the C-terminus was totally inactive, while the deletion of two LIM domains at the C-terminus partially recovered its ability to stimulate transcription. The deletion of the second LIM domain at C-terminus did not alter the activation capacity of FHL2. These results suggest that the last LIM domain at the C-terminus of FHL2 is critical for its transcription activation function, the second LIM domain at the C-terminus may be a negative regulation region, but this negative regulation depends on the last LIM domain. Mapping of transcription activation domain of FHL2 lays solid basis for further study on various FHL2 functions.","PeriodicalId":21763,"journal":{"name":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","volume":"35 7","pages":"643-8"},"PeriodicalIF":0.0,"publicationDate":"2003-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22501151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Photodynamic effects of 5-aminolevulinic acid and its hexylester on several cell lines. 5-氨基乙酰丙酸及其己酯对几种细胞系的光动力效应。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Shu-Min Wu, Qing-Guang Ren, Mi-Ou Zhou, Yi Wei, Ji-Yao Chen

{"title":"Photodynamic effects of 5-aminolevulinic acid and its hexylester on several cell lines.","authors":"Shu-Min Wu, Qing-Guang Ren, Mi-Ou Zhou, Yi Wei, Ji-Yao Chen","doi":"","DOIUrl":"","url":null,"abstract":"5-aminolevulinic acid (ALA) and its hexyl-ester (He-ALA) has shown promising results in photodynamic detection and therapy of tumors. In this work, the photodynamic effects of ALA and He-ALA on neuroblastoma cells, hepatoma cells and fibroblast cells were comparatively studied. With the detection of fluorescence emission spectra, protoporphyrin IX (PpIX) induced by ALA or He-ALA was observed in these three cell lines. Confocal laser scanning microscope showed the diffuse PpIX fluorescence in cytoplasm of neuroblastoma cells. The kinetics of PpIX accumulation were different in these three kinds of cells. The PpIX content in hepatoma cells and fibroblast cells continuously increased with the incubation time of drugs until 12 h, while in neuroblastoma cells the PpIX content saturated around 8 h after incubation with ALA or He-ALA. In addition, the PpIX concentration in neuroblastoma cells was obviously higher than that in hepatoma cells and fibroblast cells, indicating that the PpIX production is cell line dependent. When incubated with ALA and irradiated with light, near 90% neuroblastoma cells were destroyed, while for hepatoma cells and fibroblast cells the death rate was around 50%. The results demonstrate that neuroblastoma cells are more sensitive to ALA-PDT and the neuro-tumor cells may be well suited for the treatment of ALA mediated photosensitization. Comparing to ALA, He-ALA can reach the similar results concerned PpIX production and PDT damaging in all three kinds of cells but with 10 times lower incubation concentration, demonstrating that He-ALA has higher efficiency than ALA on inactivation of cancer cells in vitro.","PeriodicalId":21763,"journal":{"name":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","volume":"35 7","pages":"655-60"},"PeriodicalIF":0.0,"publicationDate":"2003-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22501582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Expression, purification, and characterization of recombinant Saccharomyces cerevisiae adenosine kinase. 重组酿酒酵母腺苷激酶的表达、纯化和鉴定。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Xiao-Bing Lu, Hai-Zhen Wu, Jiang Ye, Yi Fan, Hui-Zhan Zhang

引用次数: 0

Characterization of a new bradykinin-potentiating peptide (TmF) from Trimeresurus mucrosquamatus. 一种新的缓激肽增强肽(TmF)的鉴定。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Yong-Hong Jia, Dong-Sheng Li, Shao-Wen Zhu, Li-Yue Zhang, Li-Sheng Ding, Wan-Yu Wang, Yu-Liang Xiong

{"title":"Characterization of a new bradykinin-potentiating peptide (TmF) from Trimeresurus mucrosquamatus.","authors":"Yong-Hong Jia, Dong-Sheng Li, Shao-Wen Zhu, Li-Yue Zhang, Li-Sheng Ding, Wan-Yu Wang, Yu-Liang Xiong","doi":"","DOIUrl":"","url":null,"abstract":"A novel bradykinin-potentiating peptide (BPP), designated as TmF, has been purified to homogeneity from the venom of Trimeresurus mucrosquamatus by 70% cold methanol extraction, Sephadex G-15 gel filtration and reverse-phase high performance liquid chromatography (RP-HPLC). The amino acid sequence of TmF was determined to be pGlu-Gly-Arg-Pro-Leu-Gly-Pro-Pro-Ile-Pro-Pro (pGlu denotes pyroglutamic acid), which shared high homology with other BPPs. The molecular mass of TmF was 1.1107 kD as determinated by electrospray ionization-mass spectrometry (ESI-MS), which was in accordance with the calculated value of 1.1106 kD. The potentiating unit of TmF to bradykinin-induced (BK-induced) contraction on the guinea-pig ileum in vitro was (1.13 +/-0.3) unit (mg/L), and TmF (5.0 x10(-4) mg/kg) increased the pressure-lowering-effect of bradykinin (5.0 x10(-5 )mg/kg) with approximate descent value of (14 +/-2) mmHg. In addition, TmF inhibited the conversion of angiotensin I to angiotensin II, 2 x10(-3) mg of TmF caused 50% inhibition (IC(50)) of angiotensin- converting enzyme (ACE) hydrolyzing activity to bradykinin.","PeriodicalId":21763,"journal":{"name":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","volume":"35 7","pages":"619-23"},"PeriodicalIF":0.0,"publicationDate":"2003-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22501147","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Identification of protein spots in silver-stained two-dimensional gels based on chemically assisted fragmentation by MALDI-TOF-PSD mass spectrometry]. [基于MALDI-TOF-PSD质谱的化学辅助碎片法鉴定银染色二维凝胶中的蛋白质斑点]。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Ping Chen, Song Nie, Jin-Yun Xie, Song-Ping Liang, Chong Peng, Gui-Yuan Li

引用次数: 0

A novel polymorphism A(+884)-->G in the hepatic lipase gene and its association with coronary artery disease. 肝脂肪酶基因新多态性A(+884)- >G及其与冠状动脉疾病的关系

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Zhi-Guang Su, Si-Zhong Zhang, Li Zhang, Yu Tong, Cui-Ying Xiao, Yi-Ping Hou, Lin-Chuan Liao

{"title":"A novel polymorphism A(+884)-->G in the hepatic lipase gene and its association with coronary artery disease.","authors":"Zhi-Guang Su, Si-Zhong Zhang, Li Zhang, Yu Tong, Cui-Ying Xiao, Yi-Ping Hou, Lin-Chuan Liao","doi":"","DOIUrl":"","url":null,"abstract":"Hepatic lipase (HL) activity may influence susceptibility to coronary artery disease (CAD). Association between the single nucleotide polymorphisms (SNPs) in the HL gene with the occurrence of CAD has been investigated thoroughly, but to date most studies focused on the base variation in the promoter of HL gene, little is known about the variation in the coding region. In present study, the SNP in all exons of the HL gene were analyzed. All 9 exons with their flanking sequences of the HL gene were amplified from the Chinese patients with CAD and normal controls by PCR technique, and the PCR products were detected by denaturing high performance liquid chromatography (DHPLC) and sequenced with a dideoxy terminal termination method. As the result, a novel SNP A(+884)-->G within the sixth exon of HL gene was found, the 276 codon AAA was changed into AGA and resulted in the substitution of arginine for lysine. Compared with the control group, more CAD patients carried the G+884 allele (AG+GG) (54.9% vs. 41.5%, chi(2)=6.164, df=2, P=0.046). The prevalence of the G+884 allele was significantly higher in the CAD patients than that in control subjects (31.4% vs. 21.3%, chi(2) =4.652, df=1, P=0.031). Data from the linkage disequilibrium analysis showed that the A(+884)-->G polymorphism was strong in linkage disequilibrium with the T(-2)-->C variation we identified previously(D'=0.699, 0.742 in CAD patients and controls, respectively), and the frequency of the C(-2)/G(+884) haplotype (mutation) is significantly higher in CAD patients than that in controls (0.253 vs. 0.172, P<0.05).","PeriodicalId":21763,"journal":{"name":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","volume":"35 7","pages":"606-10"},"PeriodicalIF":0.0,"publicationDate":"2003-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22501145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Overexpression of human genes in Drosophila melanogaster by using GAL4 UAS system. 利用GAL4 UAS系统研究人类基因在黑腹果蝇中的过表达。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Xi-Zhi Ma, Li-Jun Cai, Xiao-Hui Wu, Shou-Yuan Zhao, Chang-Ben Li, Ke-Jing Deng

引用次数: 0

[Biological function of fusion protein ATF-PAI2CD]. 融合蛋白ATF-PAI2CD的生物学功能。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Xia Wang, Ping Li, Yu-Qing Zhang, Min Hou, Xing-Hui Sun, Li Tan, Yun-Song Zhu

{"title":"[Biological function of fusion protein ATF-PAI2CD].","authors":"Xia Wang, Ping Li, Yu-Qing Zhang, Min Hou, Xing-Hui Sun, Li Tan, Yun-Song Zhu","doi":"","DOIUrl":"","url":null,"abstract":"To express the fusion protein ATF-PAI2CD (urokinase-type plasminogen activator amino terminal fragment-plasminogen activator inhibitor type 2 with the region inter C and D helices deleted ) gene in E.coli and determine the biological characterization of fusion protein ATF-PAI2CD, the cDNA fragment encoding ATF-PAI2CD was cloned into the expression vector pLY-4 and transformed into E.coli JF1125. After temperature induction, the expression amount of ATF-PAI2CD account for 15% of total bacterial protein. The result was confirmed by Western blot. ATF-PAI2CD protein was isolated and purified by washing and solubilization of inclusion body, renaturation and ion exchange chromatography. The final product displayed a single band with a corresponding molecular weight 62 kD in SDS-PAGE. The purity was over 90%, the protein yield was 50% and the specific activity was 12 000 IU/mg. The PAI activity was measured by chromogenic assay. The purified fusion protein inhibited urokinase-type plasminogen activator as measured by milk-agarose plate assay, and bound to human lung cancer cells via uPA receptor (uPAR), which was confirmed by radio competition experiments. The results indicate that the biological characteristics of ATF-PAI2CD were very similar to those of the wide type PAI-2 (or mutants PAI-2, PAI-2CD) and to pro-uPA in binding to uPAR-bearing cells.","PeriodicalId":21763,"journal":{"name":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","volume":"35 7","pages":"624-8"},"PeriodicalIF":0.0,"publicationDate":"2003-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22501148","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Identification of interaction between PAI-2 and IRF-3. PAI-2与IRF-3相互作用的鉴定。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Yu-Qing Zhang, Ping Li, Min Hou, Xia Wang, Jing Fan, Li Tan, Yun-Song Zhu

引用次数: 0

Expression of recombinant human ICOS and in vitro characterization of its bioactivity on B lymphocytes. 重组人ICOS的表达及其对B淋巴细胞生物活性的体外表征。

Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica Pub Date : 2003-07-01

Zhong-Bin Deng, Chang-Ming Lu, Wei-Da Huang, Li-Qin Shen, Wei Zhu, Hong-Bing Ma, Pan-Sheng Fan, Xue-Guang Zhang

引用次数: 0