Metal ion accessibility of histidine-modified superfolder green fluorescent protein expressed in Escherichia coli

被引：15

作者：

Tansila, Natta ^{[2
]}

Becker, Kristian ^{[1
]}

Isarankura-Na-Ayudhya, Chartchalerm ^{[2
]}

Prachayasittikul, Virapong ^{[2
]}

Bulow, Leif ^{[1
]}

机构：

[1] Lund Univ, Ctr Chem & Chem Engn, Dept Pure & Appl Biochem, Lund, Sweden

[2] Mahidol Univ, Fac Med Technol, Dept Clin Microbiol, Bangkok 10700, Thailand

来源：

BIOTECHNOLOGY LETTERS | 2008年 / 30卷 / 08期

关键词：

accessible surface area; fluorescent quenching; green fluorescent protein; histidine; metal binding;

D O I：

10.1007/s10529-008-9692-7

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Green fluorescent protein (GFP) is frequently utilized for metal ion detection and quantification. To improve the metal binding potential of GFP, three residues (N146, F165, and L201) were substituted to histidines. Each variant responded differently upon interaction with metal ions. More than 80% of N146H, having the most accessible surface area, could bind to immobilized metal ions. However, only F165H exhibited significant differences in quenching by soluble metal ions (22% fluorescence decrease) in comparison with the template protein (12%). These findings can be utilized for designing GFP variants for metal binding and sensor applications.

引用

页码：1391 / 1396

页数：6

共 15 条

[11] Interaction analysis of chimeric metal-binding green fluorescent protein and artificial solid-supported lipid membrane by quartz crystal microbalance and atomic force microscopy [J].