M. Diaz et al., Characterization of intestinal Na+-K+-ATPase in the gilthead seabream (Sparus aurata L.). Evidence for a tissue-specific heterogeneity, COMP BIOC B, 121(1), 1998, pp. 65-76
Citations number
36
Categorie Soggetti
Biochemistry & Biophysics
Journal title
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY
Gilthead seabream intestine contains both a Mg2+ -dependent Na+-K+-ATPase w
hich is completely inhibited by 1 x 10(-3) M ouabain, and also a residue-AT
Pase activity that is entirely ouabain-insensitive. The maximal activity of
intestinal Na+-K+-ATPase (35.15 mu mol inorganic phosphate (P-i) mg protei
n(-1) h(-1)) was observed in the microsomal fraction at 35 degrees C, pH 7.
5, 2-5 mM MgCl2, 5 mM ATP, 10 mM K+ and 200 mM Na+. The intestinal Na+-K+-A
TPase of gilthead seabream exhibits similar characteristics to other teleos
t Na+-K+-ATPases regarding pH dependence, Mg2+/ATP optimal ratios and ouaba
in sensitivity, but exhibits unusual sensitivity to ionic strength and cati
on promoted cooperative activation and higher affinity for magnesium and AT
P when compared to other marine and euryhaline teleosts. The Arrhenius plot
for intestinal Na+-K+-ATPase showed a break point at 15.41 degrees C, with
similar activation energies above and below the discontinuity point. The a
nalysis of polar lipid fatty acid composition was correlated to the break p
oint in the Arrhenius plot, suggesting a regulatory role of the lipid micro
environment on the enzyme activity. Finally, the kinetic characteristics, t
emperature-activity relationship, fatty acid composition and substrate depe
ndence of intestinal Na+-K+-ATPase are compared with literature data and di
scussed on the basis of the differences between species and also between os
moregulatory tissues. It is concluded that the Na+-K+-ATPase of Sparus aura
ta is differently and specifically expressed between the osmoregulatory org
ans. (C) 1998 Elsevier Science Inc. All rights reserved.