[Effect of the medium on functional and structural properties of serum albumins. IV. The state of human serum albumin in the pH range from 5 to 10].
In order to investigate the effects of temperature and ionic strength on the N-B-transition and the alkaline denaturation of the human serum albumin, the pH-dependences of fluorescence position and relative yield of Trp-24 and of protein bound dye ANS were measured. The measurements were carried out at temperatures from 10 to 45 degrees C and ionic strengths (NaCl) from 0.001 to 0.2. The pH-induced structural transitions have different realization in environments of tryptophanyl and tightly bound ANS. The alkaline denaturation does not change the Trp-214 fluorescence. The N-B-transition gives rise to the slight polarity and/or mobility lowering in the Trp-214 environment (the shorter-wave-length spectral shift). Increase in the temperature and ionic strength induces the shift of the transition midpoint from ca. 8 to 8.7 and reduces the spectral shift amplitude. At low ionic strengths, the new structural transition in the Trp-214 environment is observed at pH change from 6.7 to 5.7. This transition is not observable using ANS fluorescence. The N-B-transition is accompanied by an enhancement and longer-wavelength shift of the ANS fluorescence spectra. The transition midpoint is independent of temperature, but is shifted to lower pH values at a decrease of ionic strength value. At ionic strengths less than or equal to 0.01 the shorter-wavelength spectral shift is seen at pH from 7.5 to 9, which seems to reflect the disulfide B-A-isomerisation. The alkaline denaturation gives rise to the sharp quenching of ANS fluorescence, probably due to the ANS binding site decomposition.(ABSTRACT TRUNCATED AT 250 WORDS)
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D012709:Serum Albumin / Q000032:analysis*; D013050:Spectrometry, Fluorescence
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D012709:Serum Albumin
Vliianie sredy na funktsional'nye i strukturnye svoĭstva syvorotochnykh al'buminov. IV. Sostoianie syvorotochnogo al'bumina cheloveka v zone pH ot 5 do 10.