集胞藻类金属硫蛋白的纯化、性质和溶液构象的研究

CORC > 北京大学 > 生命科学学院

	集胞藻类金属硫蛋白的纯化、性质和溶液构象的研究; Studies on the purification, characterization and conformation in solution of metallothionein-like substance from Synechocystis 6803
	陈正佳 ; 李晓凡 ; 施定基 ; 李玲媛 ; 茹炳根
刊名	植物学报
	1999
关键词	集胞藻类金属硫蛋白分离和纯化性质溶液构象 Synechocystis sp. PCC 6803 metallothionein-like compound isolation and purification characterization conformation in solution
英文摘要	集胞藻Synechocysitissp.PCC6803光照培养，加入100μmol/LZnCl2诱导藻内类金属硫蛋白（MT-like）的表达。离心收集鲜藻，超声波破碎细胞，离心除沉淀。上清液72℃加热3min去杂蛋白，离心，上清液依次经过分子筛层析柱，阴离子交换柱和脱盐柱，可得蓝藻类金属硫蛋白。5L培养液收集鲜藻7.6g，一次上样1．52g鲜藻的裂解物，冻干后得15mg纯化的蛋白样，为鲜藻重的0．1％．经分析其等电点为pH4．5，分子量为6．986kD，由58个氨基酸残基组成。其序列中含有较多疏水氨基酸残基（36％），但其半胱氨酸含量仅为5％。CD（圆二色性）图谱表明其二级结构主要为无规卷曲，...; The cyanobacterium Synechocystis sp. PCC 6803 was cultivated for 5 days in sterile liquid medium BG-11 at room temperature under fluorescent light, aerated by stirring under sterile condition. ZnCl2 was then added into the medium at final concentration of 50 mu mol/L to induce the expression of the metallothionein-like (MT-like) compound. 5 days later, the ZnCl2 concentration was increased to 100 mu mol/L. After 2 days, cells were collected by centrifugation, then disrupted by ultrasonic wave. After centrifugation, the supernatant was treated at 72 degrees C for 3 min to denature the unstable proteins. Again after centrifugation, the supernatant was applied to Sephacryl S-100 molecular sieve column, then DEAE-Sepharose F. F. ion exchange column and Sephadex G-25 column for desalination. The Zn-binding fraction was frozen dried. In 5 L cultures, 7.6 g wet cyanobacterial cells H-ere obtained. Each time the lysate of 1.52 g of them was processed, finally 1.5 mg purified MT-like compound was obtained, which was 0.1% of the total weight of wet cyanobacterial cells. Isoelectric focusing (IEF) showed that its pi was pH 4.5. Mass spectrometry and amino acid composition analysis indicated that the molecular weight of the protein was 6.986 kD. Amino acid composition analysis shelved that the content of hydrophobic amino acid residues was 36%, but the Cys content was not very high (only 5%). Ultraviolet absorption spectra showed that Zn-MT-like binding complex also hsd a high absorbance at 220 nm. CD spectroscopy proved that the secondary structure of this protein was a random coil,and that there were no alpha-helix and beta-sheet in the structure. FT-IR spectroscopy of cyanobacterial MT-like compound was similar to that of the typical MT of mammal.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000079244300008&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; SCI(E); 中文核心期刊要目总览(PKU); 中国科学引文数据库(CSCD); 2; 02; 150-155; 41
语种	中文
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/320219]
专题	生命科学学院
推荐引用方式 GB/T 7714	陈正佳,李晓凡,施定基,等. 集胞藻类金属硫蛋白的纯化、性质和溶液构象的研究, Studies on the purification, characterization and conformation in solution of metallothionein-like substance from Synechocystis 6803[J]. 植物学报,1999.
APA	陈正佳,李晓凡,施定基,李玲媛,&茹炳根.(1999).集胞藻类金属硫蛋白的纯化、性质和溶液构象的研究.植物学报.
MLA	陈正佳,et al."集胞藻类金属硫蛋白的纯化、性质和溶液构象的研究".植物学报 (1999).