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核苷二磷酸激酶A的异构及其分子机制 被引量:3
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作者 熊盛 钱垂文 +6 位作者 王一飞 黄立 李久香 严玖凤 王小宁 张晓伟 毕志刚 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2005年第6期815-821,共7页
对核苷二磷酸激酶A(NDPKA)的异构及其分子机制进行研究.还原和非还原SDSPAGE观察重组人核苷二磷酸激酶A(rhNDPKA)的异构;RPHPLC分析rhNDPKA异构体的反相色谱行为,并测定rhNDPKA异构体的酶活性;多角度激光散射法测定rhNDPKA异构体在溶液... 对核苷二磷酸激酶A(NDPKA)的异构及其分子机制进行研究.还原和非还原SDSPAGE观察重组人核苷二磷酸激酶A(rhNDPKA)的异构;RPHPLC分析rhNDPKA异构体的反相色谱行为,并测定rhNDPKA异构体的酶活性;多角度激光散射法测定rhNDPKA异构体在溶液中的表观分子量;飞行质谱分析异构体的质量肽谱.结果发现,rhNDPKA在非还原SDSPAGE上表现为4条带,对应于NDPKA的氧化型、还原型、氧化型二聚体和还原型二聚体,其分子量分别为18.1kD、21.3kD、35.2kD和38.3kD.RPHPLC发现,还原型rhNDPKA和氧化型rhNDPKA疏水性有差异.新鲜制备的rhNDPKA在纯水溶液中,经空气氧化后,逐渐由还原型向氧化型过渡,而还原剂或生理盐水可使rhNDPKA稳定于还原型或氧化型.酶活测定结果表明,还原型rhNDPKA比活性为1965±166Umg,氧化型rhNDPKA比活性为974±53Umg.多角度激光散射检测发现,还原型rhNDPKA在溶液中仍可形成六聚体.质量肽谱结果证明,在氧化型rhNDPKA中,C4和C145位巯基形成二硫键,而C109位巯基游离存在.根据本文所确定的NDPKA单体中的二硫键位置,推导出rhNDPKA单体异构体和二聚体异构体的变构原理,这为进一步研究NDPKA的多能性调节机制打下了良好基础. 展开更多
关键词 核苷二磷酸激酶A 异构 多角度激光散射 飞行质谱 二硫键 分子机制
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Nm23/NDPK的多种生物活性及其机制研究进展 被引量:4
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作者 熊盛 邢少璟 +1 位作者 钱垂文 王一飞 《中国病理生理杂志》 CAS CSCD 北大核心 2006年第10期2066-2070,共5页
Human Nm23 family consists of eight gene products that have been implicated in cellular differentiation,development and apoptosis,as well as oncogenesis and tumor metastasis.The protein products of Nm23 are nucleoside... Human Nm23 family consists of eight gene products that have been implicated in cellular differentiation,development and apoptosis,as well as oncogenesis and tumor metastasis.The protein products of Nm23 are nucleoside diphosphate kinases(NDPKs),the key metabolic enzymes that catalyze the synthesis of nucleoside triphosphates(NTP) by transfer of the terminal phosphate between NDP and NTP.Recent investigations are focused on the extraordinary pleiotropy and its mechanism of Nm23/NDPK.In this article,we review the recent progress in studies of the mechanism of Nm23/NDPK as metastasis suppressors,and other bioactivities of NDPK out of phosphate transfer enzyme,as well as the character of NDPK’s quaternary structure and the relation between its structure and function.In addition to these,the potential applications of NDPK as an enhancer of antiviral drugs or Nm23 as a drug target were also described.Findings herein summarized provide new and intriguing suggestions for a more extensive understanding of the biological functions of the star molecule,Nm23/NDPK. 展开更多
关键词 基因 Nm23 核苷二磷酸激酶
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