Non-covalent interaction of 2', 4', 5', 7'-tetrabromo-4, 5, 6, 7-tetrachlorofluorescein with proteins and its application.
The interactions of 2', 4', 5', 7'-tetrabromo-4, 5, 6, 7-tetrachlorofluorescein (TBTCF) with BSA, ovalbumin (OVA) and poly-L-lysine (PLYS) at pH 3.70 have been investigated by combination of the spectral correction technique and the Langmuir isothermal adsorption. The active connection actions such as ion pairs, van der Waals' force, hydrogen bond, hydrophobic bond were proposed to explain the non-covalent interaction between TBTCF and BSA, OVA and PLYS. Effects of the electrolyte and high temperature indicated that union of the active connections between TBTCF and BSA and OVA was too firm to be destroyed. The relationship between the binding number of TBTCF and variety fraction of the amino acid residues was analyzed. The binding number of TBTCF depended on the number of positively charged amino acid residues. The other amino acid residues surrounded and seized TBTCF by hydrogen bonds and hydrophobic bonds when the electrostatic attraction pulled TBTCF to link protein. In addition, a novel method named the absorbance ratio difference was established for determination of protein in trace level and was applied with much higher sensitivity than the ordinary method.
Gao
Hong-Wen
HW
State Key Laboratory of Pollution Control and Resource Reuse, College of Environmental Science and Engineering, Tongji University, Shanghai, P R China. hwgao@mail.tongji.edu.cn
Zhao
Jian-Fu
JF
Yang
Qiao-Zhi
QZ
Liu
Xiang-Hu
XH
Chen
Ling
L
Pan
Lu-Ting
LT
D000818:Animals; D002417:Cattle; D002645:Chickens; D005452:Fluoresceins / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D010047:Ovalbumin / Q000737:chemistry / Q000378:metabolism; D025941:Protein Interaction Mapping; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D013053:Spectrophotometry; D055672:Static Electricity; D013696:Temperature
C099822:4'-bromo-4,5,6,7-tetrachlorofluorescein; D005452:Fluoresceins; D008024:Ligands; D012710:Serum Albumin, Bovine; D010047:Ovalbumin