Use of fluorescence enhancement technique to study bilirubin-albumin interaction

被引:48
作者
Athar, H [1 ]
Ahmad, N
Tayyab, S
Qasim, MA
机构
[1] Aligarh Muslim Univ, JN Med Coll, Dept Biochem, Aligarh 202002, Uttar Pradesh, India
[2] Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
关键词
fluorescence enhancement; serum albumin; bilirubin binding;
D O I
10.1016/S0141-8130(99)00056-2
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Bilirubin-albumin solution gave an emission spectrum in the wavelength range 500-600 nm with emission maxima at 528 nm when excited at 487 nm. The magnitude of fluorescence intensity increased on increasing bilirubin/albumin molar ratio. At three different albumin concentrations, namely, 1.0, 2.5 and 10.0 mu M, there was an initial linear increase in fluorescence up to a molar ratio 1.0 in all cases beyond which it sloped off or decreased. This fluorescence enhancement was used to calculate the binding parameters of bilirubin-albumin interaction and the value of binding constant was found to be 1.72 x 10(7) l/mol similar to the published values obtained with other methods. Different serum albumins, namely, human (HSA), goat (GSA), pig (PSA) and dog serum albumins (DSA) bound bilirubin with almost the same affinity when studied by the technique of fluorescence enhancement. Bilirubin-albumin interaction was also studied at different pH and ionic strengths. There was a decrease in bilirubin-albumin complex formation on either decreasing the pH from 9.0 to 7.0 or increasing the ionic strength from 0.15 to 1.0. These results suggest that the technique of fluorescence enhancement can be used successfully to study the bilirubin-albumin interaction. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:353 / 358
页数:6
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