Biophysical studies on the correction of uremic human serum albumin binding defects by in vitro charcoal adsorption treatment.
Spectrofluorimetry, flow microcalorimetry, and differential scanning microcalorimetry (DSMC) were used to study the conformation, binding function, and ligand loading of uremic albumin obtained from the blood plasma of 2 end-stage renal disease (ESRD) patients before and after charcoal plasma treatment at different pH values (3.0-9.0). The spectrofluorimetric patterns of conformational N-F transition at low pH (4.2-3.5) are practically identical for both samples of uremic human serum albumin (HSA) and control HSA from healthy donors. After the charcoal treatment at pH 3.0 and 4.0, the enthalpies of complexing on uremic HSA with bromsulfalein and sodium dodecylsulfate approach that of donor HSA. The binding affinity of uremic HSA for sodium octanoate, phenol red, and salicylic acid following low pH charcoal treatment even exceed those of donor HSA. At the same time the charcoal treatment of uremic plasma at neutral and alkaline pH does not notably improve the binding characteristics of isolated HSA. Adsorption at low pH values completely restores the tryptophan fluorescence spectrum position of uremic albumin and improves the thermodynamic characteristics of its melting process. Using DSMC data, it can nevertheless be concluded that some conformational changes or a certain amount of high-affinity bound endogenous ligands still remain after low pH uremic HSA purification. The latter conclusion requires additional improvements of adsorption treatment of uremic plasma.
Nikolaev
V
V
Institute of Experimental Pathology, Oncology and Radiobiology, Academy of Sciences, Kiev, Ukraine.
Sarnatskaya
V
V
von Appen
K
K
Ivanov
A
A
Rotellar
E
E
Haspar
M
M
Klinkmann
H
H
D000327:Adsorption; D000328:Adult; D001665:Binding Sites; D002152:Calorimetry, Differential Scanning; D002606:Charcoal / Q000008:administration & dosage / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007676:Kidney Failure, Chronic / Q000097:blood / Q000188:drug therapy* / Q000503:physiopathology; D008297:Male; D008875:Middle Aged; D011485:Protein Binding; D011489:Protein Denaturation / Q000187:drug effects; D012015:Reference Standards; D012709:Serum Albumin / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D016896:Treatment Outcome; D014511:Uremia / Q000097:blood / Q000188:drug therapy* / Q000503:physiopathology
D012709:Serum Albumin; D002606:Charcoal