Anion-induced refolding of human serum albumin under low pH conditions.
We studied the effect of various anions (of acids and salts) on the acid denatured state of HSA by near-UV circular dichroism (CD), far-UV CD, 1-anilinonaphthalene-8-sulfonate (ANS) binding, tryptophan fluorescence and thermal transition. Addition of different acids and salts caused an induction of alpha-helical structure as evident from the increase in the mean residue ellipticity (MRE) value at 222 nm and loss of ANS binding sites exhibited by the decrease in the ANS fluorescence intensity at 480 nm. However, the concentration range of acids/salts required to bring about the transition varied greatly among different acids and salts. Among various acids/salts tested, K(3)Fe(CN)(6) was found to be most effective whereas HCl and KCl were least effective in inducing the properties close to native structure. Further, they followed the electroselectivity series. The near-UV CD spectra showed an increase in MRE towards the native state, whereas the tryptophan fluorescence emission spectra produced a red shift of about 6 nm on addition of KClO(4). The temperature-induced transition in the presence of 40 mM KClO(4) monitored by ellipticity measurements at 222 nm was characterized by the presence of an intermediate state in the temperature range 30-50 degrees C having abundant secondary structure. These results suggest that human serum albumin at low pH and in the presence of acids or salts exists in a partially folded state characterized by native-like secondary structure and tertiary folds.
Muzammil
S
S
Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, India.
Kumar
Y
Y
Tayyab
S
S
D000817:Anilino Naphthalenesulfonates; D000838:Anions / Q000737:chemistry; D002942:Circular Dichroism; D005453:Fluorescence; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012709:Serum Albumin / Q000737:chemistry*
C050774:2-anilinonaphthalene-8-sulfonic acid; D000817:Anilino Naphthalenesulfonates; D000838:Anions; D012709:Serum Albumin