Cation binding to parvalbumin studied by 113Cd and 23Na NMR. Peak assignment of rabbit (pI 5.5) parvalbumin.

Abstract

Cation binding to three apoparvalbumins was studied by means of 113Cd NMR. The 3 parvalbumins that were investigated were carp pI 4.25, rabbit pI 5.5 and pike pI 5.0. The results showed that Cd2+ ions bind to the EF and CD sites of carp apoparvalbumin pI 4.25 with about the same affinity. For rabbit (pI 5.5) apoparvalbumin, Cd2+ binds preferentially to the EF site, while for pike (pI 5.0) apoparvalbumin, it was the CD site that exhibited somewhat higher affinity for Cd2+. The effect of Mn2+ on the 113Cd signals of rabbit parvalbumin was used to assign the 113Cd NMR signals to the EF and CD sites. The Mn2+ paramagnetic effect on rabbit and pike parvalbumins differed from that obtained for carp parvalbumin. This is in agreement with the assumption that the beta-lineage parvalbumins possess a third external site of higher affinity than the alpha-lineage parvalbumins. Furthermore, 23Na NMR was used to study Na+-Mg2+ competition in the native carp (pI 4.25) parvalbumin. The results showed that Na+ and Mg2+ compete for the same site, the third external site.