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郑晓峰, Chuan-Yun Li, *, Quan Yu, Zhi-Qiang Ye, Ying Sun, Quanyuan He, , Xiao-Mo Li, Wuxue Zhang, Jingchu Luo, Xiaocheng Gu, Xiaofeng Zheng, Liping Wei
Cell Research (2007)17: 357-362,-0001,():
-1年11月30日
The use of oseltamivir, widely stockpiled as one of the drugs for use in a possible avian influenza pandemic, has been reported to be associated with neuropsychiatric disorders and severe skin reactions, primarily in Japan. Here we identified a nonsynonymous SNP (single nucleotide polymorphism) in dbSNP database, R41Q, near the enzymatic active site of human cytosolic sialidase, a homologue of virus neuraminidase that is the target of oseltamivir. This SNP occurred in 9.29% of Asian population and none of European and African American population. Our structural analyses and Ki measurements using in vitro sialidase assays indicated that this SNP could increase the unintended binding affinity of human sialidase to oseltamivir carboxylate, the active form of oseltamivir, thus reducing sialidase activity. In addition, this SNP itself results in an enzyme with an intrinsically lower sialidase activity, as shown by its increased Km and decreased Vmax values. Theoretically administration of oseltamivir to people with this SNP might further reduce their sialidase activity. We note the similarity between the reported neuropsychiatric side effects of oseltamivir and the known symptoms of human sialidase-related disorders. We propose that this Asian-enriched sialidase variation caused by the SNP, likely in homozygous form, may be associated with certain severe adverse reactions to oseltamivir.
Asia,, SNP,, neuraminidase inhibitor,, oseltamivir,, sialidase,, bioinformatics
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郑晓峰, Yamei Yu, Lanfen Li, Xiaofeng Zheng, Yu-He Liang, Xiao-Dong Su *
Biochimica et Biophysica Acta 1764(2006)153-156,-0001,():
-1年11月30日
Imidazolonepropionase (EC 3.5.2.7) is the third enzyme of the histidine degradation pathway that has been conserved from bacteria to eukaryotes. The enzyme is the only one with unknown three-dimensional structure in this pathway. In this work, Bacillus subtilis imidazolonepropionase (HutI) was expressed in E. coli and purified to homogeneity. After thrombin digestion, high quality crystals were obtained by hanging-drop vapor diffusion method. The best crystal diffracted to 2.0 A° and belonged to the space group P21 with unit-cell parameters a=57.73A°, b=106.34A°, c=66.47A°, b=89.93.
Imidazolonepropionase, Crystallization, Thrombin-digestion, X-ray diffraction
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【期刊论文】Serum levels of preS antigen (HBpreSAg) in chronic hepatitis B virus infected patients
郑晓峰, Min Lian†, , Xu Zhou†, Lai Wei†, Shihong Qiu, Tong Zhou, Lanfen Li, Xiaocheng Gu, Ming Luo, and Xiaofeng Zheng*
Virology Journal 2007, 4: 93,-0001,():
-1年11月30日
Background: Hepatitis B virus (HBV) infection is a serious health problem worldwide. Treatment recommendation and response are mainly indicated by viral load, e antigen (HBeAg) seroconversion, and ALT levels. The S antigen (HBsAg) seroconversion is much less frequent. Since HBeAg can be negative in the presence of high viral replication, preS antigen (HBpreSAg) might be a useful indicator in management of chronic HBV infection.Results: A new assay of double antibody sandwich ELISA was established to detect preS antigens. Sera of 104 HBeAg-negative and 50 HBeAg-positive chronic hepatitis B patients have been studied and 23 HBeAg-positive patients were enrolled in a treatment follow-up study. 70% of the HbeAgpositive patients and 47% of the HBeAg-negative patients showed HBpreSAg positive. Particularly, in the HBeAg-negative patients, 30 out of 47 HBpreSAg positive patients showed no evidence of viral replication based on HBV DNA copies. A comparison with HBV DNA copies demonstrated that the overall accuracy of the HBpreSAg test could reach 72% for active HBV replication. HBpreSAg changes were well correlated with changes of HBsAg, HBV DNA and ALT levels during the course of IFN-α treatment and follow-up. HBeAg positive patients responded well to treatment when reduction of HBpreSAg levels was more pronounced.Conclusion: Our results suggested that HBpreSAg could be detected effectively, and well correlated with HBsAg and HBV DNA copies. The reduction of HBpreSAg levels in conjunction with the HBV DNA copies appears to be an improved predictor of treatment outcome.
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郑晓峰, Xiaofeng Zheng and Philip C. Bevilacqua*
Nucleic Acids Research, 2001, Vol. 29, No.2 e6,-0001,():
-1年11月30日
We have developed a semi-synthetic approach for preparing long stretches of DNA (>100 bp) containing internal chemical modifications and/or non-Watson-Crick structural motifs which relies on splint-free, cell-free DNA ligations and recycling of side-products by non-PCR thermal cycling. A double-stranded DNA PCR fragment containing a polylinker in its middle is digested with two restriction enzymes and a small insert (<20 bp) containing the modification or non-Watson-Crick motif of interest is introduced into the middle. Incorrect products are recycled to starting materials by digestion with appropriate restriction enzymes, while the correct product is resistant to digestion since it does not contain these restriction sites. This semi-synthetic approach offers several advantages over DNA splint-mediated ligations, including fewer steps, substantially higher yields (<60%overall yield) and ease of use. Thismethod has numerous potential applications, including the introduction of modifications such as fluorophores and cross-linking agents into DNA, controlling the shape of DNA on a large scale and the study of nonsequence-specific nucleic acid-protein interactions.
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【期刊论文】Systematic high-yield production of human secreted proteins in Escherichia coli
郑晓峰
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-1年11月30日
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