Metal ion accessibility of histidine-modified superfolder green fluorescent protein expressed in Escherichia coli.
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.
Tansila
Natta
N
Department of Clinical Microbiology, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand.
Becker
Kristian
K
Isarankura Na-Ayudhya
Chartchalerm
C
Prachayasittikul
Virapong
V
Bülow
Leif
L
D004926:Escherichia coli / Q000378:metabolism*; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000378:metabolism*; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013499:Surface Properties
D008670:Metals; D050505:Mutant Proteins; D049452:Green Fluorescent Proteins; D006639:Histidine