Hydrophobic interaction of alcian blue with soluble and erythrocyte membrane proteins.
Alcian Blue (AB), a cationic dye widely employed for monitoring negative surface charge variations on red blood cell (RBC), platelet and glomerular membranes of patients with nephrotic syndromes, was found in fact to aggregate with itself and precipitate in the pH range 7.0-7.8, i.e., at the physiological pH values used for performing the binding assay between the dye and cell surfaces. This aggregation appears to be essentially hydrophobic as it is insensitive to urea but fully prevented in presence of 2% zwitterionic detergent. In addition, AB binds to most RBC membrane proteins solubilized by urea-detergent extraction, again suggesting hydrophobic interaction. AB also interacts with freely soluble proteins such as haemoglobin and myoglobin; such binding is disrupted by ethylurea and/or 2% zwitterionic detergent, typical inhibitors of hydrophobic liaisons. AB also strongly binds to myoglobin with all the negative charges blocked by esterification of the carboxyl groups, again ruling out direct interaction via surface negative charges. It is concluded that AB binding to the RBC surface can hardly monitor variations in surface charge due to sialic acid residues but, at best, variations in surface hydrophobicity.
Ghiggeri
G M
GM
Nephrology Section, Hospital of Lavagna, Italy.
Candiano
G
G
Ginevri
F
F
Mutti
A
A
Bergamaschi
E
E
Alinovi
R
R
Righetti
P G
PG
D000328:Adult; D000423:Alcian Blue / Q000032:analysis*; D001798:Blood Proteins / Q000032:analysis*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000032:analysis*; D006454:Hemoglobins / Q000032:analysis; D006801:Humans; D007211:Indoles / Q000032:analysis*; D009211:Myoglobin / Q000032:analysis
D001798:Blood Proteins; D006454:Hemoglobins; D007211:Indoles; D009211:Myoglobin; D000423:Alcian Blue