Oxygen-dependent oxidation of Fe(II) to Fe(III) and interaction of Fe(III) with bovine serum albumin, leading to a hysteretic effect on the fluorescence of bovine serum albumin.
The serum albumin is the most abundant protein in blood plasma and the iron is essential for many cellular processes. However, the interaction between Fe(3+) and haem-free serum albumin remains unclear. Here we provide evidence for the fact that haem-free BSA possesses one specific Fe(3+)-binding site. The binding of Fe(3+) to BSA results in a significant quenching of the Trp fluorescence of BSA. The average apparent dissociation constant value for the interaction of Fe(3+) and BSA is 3.46 x 10(-8)+/-3 x 10(-10) M at 37 degrees C and 3.30 x 10(-8)+/-5 x 10(-10) M at 25 degrees C, respectively, as determined by fluorescence titration. Addition of 50 microM Fe(2+) to 1 microM BSA results in an obvious hysteretic effect on the fluorescence of BSA. The time-dependent fluorescence quenching of BSA by Fe(2+) is not caused by the Fe(2+)-induced conformational change of BSA, but the oxygen-dependent oxidation of Fe(2+) to Fe(3+). Fe(2+) undergoes an oxygen-dependent oxidation to Fe(3+) under aerobic conditions, which is accelerated by the interaction of BSA with Fe(3+) and extensively inhibited under anaerobic conditions. The results suggest that BSA may take part in non-transferrin bound iron transfer.
Xu
Xiaolong
X
Department of Chemistry, University of Science and Technology of China, Hefei 230026, People's Republic of China. xuxl@ustc.edu.cn
Zhang
Liyun
L
Shen
Dengke
D
Wu
Hao
H
Liu
Qingliang
Q
D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D005290:Ferric Compounds / Q000737:chemistry* / Q000378:metabolism; D005296:Ferrous Compounds / Q000737:chemistry* / Q000378:metabolism; D005453:Fluorescence; D010084:Oxidation-Reduction / Q000187:drug effects*; D010100:Oxygen / Q000737:chemistry* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry
D005290:Ferric Compounds; D005296:Ferrous Compounds; D012710:Serum Albumin, Bovine; D014364:Tryptophan; D010100:Oxygen