Species differences of serum albumins: III. Analysis of structural characteristics and ligand binding properties during N-B transitions.
OBJECTIVE
The aim of this study was to investigate the characteristics of the structural transitions and changes in ligand binding properties of different albumins during the pH-dependent structural transition, often referred to as the N-B transition.


METHODS
Structural transitions were evaluated by means of spectrometry, differential scanning calorimetry and chemical modification. In addition, ligand binding properties were investigated using typical site-specific bound drugs (warfarin, phenylbutazone, ibuprofen and diazepam).


RESULTS
Conformational changes, including N-B transition, clearly occurred in albumins from all species used in this study. The conformational stabilities of all the albumins were clearly lost in the weakly alkaline pH range. This was probably the result of the destruction of salt bridges between domain I and domain III in the albumin molecule. In addition, the profiles of the ANS-induced fluorescence were different and could be classified into two patterns, suggesting that hydrophobic pockets in the albumin molecules were different for the different species. The data suggest that the amino acid residues responsible for the transitions were some of the His residues located in domain I. Further, the ligand binding properties of the albumins were slightly different but statistically significant.


CONCLUSIONS
The overall mechanisms of the N-B transition may be similar for all the albumins, but its impact is considerably different among the species in terms of both structural characteristics and ligand binding properties. Furthermore, the transitions appear to be multi-step transitions.
Kosa
T
T
Faculty of Pharmaceutical Sciences, Kumamoto University, Japan.
Maruyama
T
T
Sakai
N
N
Yonemura
N
N
Yahara
S
S
Otagiri
M
M
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D003975:Diazepam / Q000378:metabolism; D004285:Dogs; D006639:Histidine / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007052:Ibuprofen / Q000378:metabolism; D008024:Ligands; D010653:Phenylbutazone / Q000378:metabolism; D011487:Protein Conformation; D011817:Rabbits; D051381:Rats; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D013045:Species Specificity; D014859:Warfarin / Q000378:metabolism
D008024:Ligands; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D006639:Histidine; D014859:Warfarin; D010653:Phenylbutazone; D003975:Diazepam; D007052:Ibuprofen