Na+,K(+)-ATPase of human placenta during gestational hypertension: a biochemical-biophysical study.
1. Na+,K(+)-ATPase is the membrane enzyme catalysing the active transport of Na+ and K+ across the plasma membrane of animal cells. A reduced activity of Na+,K(+)-ATPase has been described in gestational hypertension in a variety of cell types, in agreement with the hypothesis that gestational hypertension can induce membrane transport modifications similar to those reported for essential hypertension. The causes of the reduced Na+,K(+)-ATPase activity are still debated. 2. The aim of the present work was to investigate the molecular mechanism of the reduced enzymic activity in gestational hypertension using as a model Na+,K(+)-ATPase purified from human placenta. Na+,K(+)-ATPase obtained from term placentas of eight healthy pregnant women and eight age-matched women with gestational hypertension was purified as previously described. 3. We observed in gestational hypertension: (i) a significant increase in the activation energies above transition temperature; (ii) a significant decrease in the fluorescence polarization of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (i.e. increased fluidity) and an increase in the mean lifetime (modified hydrophobicity); (iii) a lower Kq, suggesting an enzymic structural modification; and (iv) an increased mean lifetime and rotational relaxation time of pyrene isothiocyanate, indicating a modified ATP binding site.
Rabini
R A
RA
Department of Diabetology, INRCA Hospital, Ancona, Italy.
Zolese
G
G
Staffolani
R
R
Lucarelli
G
G
Amler
E
E
Cester
N
N
Mazzanti
L
L
D000328:Adult; D055592:Biophysical Phenomena; D001703:Biophysics; D005260:Female; D005454:Fluorescence Polarization; D006801:Humans; D006973:Hypertension / Q000201:enzymology*; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D011249:Pregnancy Complications, Cardiovascular / Q000201:enzymology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D013696:Temperature; D014364:Tryptophan
D014364:Tryptophan; D000254:Sodium-Potassium-Exchanging ATPase