Human serum albumin coordinates Cu(II) at its N-terminal binding site with 1 pM affinity.
The conditional stability constant at pH 7.4 for Cu(II) binding at the N-terminal site (NTS) of human serum albumin (HSA) was determined directly by competitive UV-vis spectroscopy titrations using nitrilotriacetic acid (NTA) as the competitor in 100 mM NaCl and 100 mM N-(2-hydroxyethyl)piperazine-N'-ethanesulfonic acid (Hepes). The log Kc (NTS) value of 12.0 +/- 0.1 was determined for HSA dissolved in 100 mM NaCl. A false log log Kc (NTS) (c) value of 11.4 +/- 0.1 was obtained in the 100 mM Hepes buffer, owing to the formation of a ternary Cu(NTA)(Hepes) complex. The impact of the picomolar affinity of HSA for Cu(II) on the availability of these ions in neurodegenerative disorders is briefly discussed.
Rózga
Małgorzata
M
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
Sokołowska
Magdalena
M
Protas
Anna Maria
AM
Bal
Wojciech
W
D001667:Binding, Competitive; D003300:Copper / Q000378:metabolism*; D006801:Humans; D019636:Neurodegenerative Diseases; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D017186:Titrimetry
D012709:Serum Albumin; D003300:Copper