The buffering power of plasma in brown bullhead ( Ameiurus nebulosus)

Abstract

Brown bullhead ( Ameiurus nebulosus) blood plasma was found to exhibit an unusually high non-bicarbonate buffer capacity (β) in relation to that of other teleost fish. In brown bullhead, the non-bicarbonate buffer capacity of plasma (β plasma), at −5.72±0.34 mmol l −1 pH unit −1 (mean±S.E.M., N=30), constituted 37% of whole blood β and was 2.5 times higher than β plasma in rainbow trout (−2.33±0.42 mmol l −1 pH unit −1; N=7). The strong buffering power of bullhead plasma was not the result of unusually high plasma protein levels. Size separation chromatography in conjunction with a spectrophotometric assay for buffering capacity were used to isolate a plasma fraction of high buffering power. SDS-polyacrylamide gel electrophoresis revealed that this fraction contained four proteins, but was dominated by a protein of approximately 68–70 kDa molecular mass. On the basis of the amino acid composition of this fraction, the dominant protein was identified as albumin. In comparison to other fish albumins, bullhead albumin appears to be histidine-rich (6.7%). Thus, the unusually high non-bicarbonate buffer capacity of bullhead plasma appears to stem from the presence in the plasma of a histidine-rich albumin.