These tools will no longer be maintained as of December 31, 2024. Archived website can be found here. PubMed4Hh GitHub repository can be found here. Contact NLM Customer Service if you have questions.


Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

  • Title: K+-dependence of electrogenic transport by the NaK-ATPase.
    Author: Gropp T, Cornelius F, Fendler K.
    Journal: Biochim Biophys Acta; 1998 Jan 19; 1368(2):184-200. PubMed ID: 9459597.
    Abstract:
    Charge translocation by the NaK-ATPase from shark rectal gland was measured by adsorption of proteoliposomes to a planar lipid membrane. The proteoliposomes were prepared by reconstitution of purified NaK-ATPase into liposomes consisting of E. coli lipids. The protein was activated by applying an ATP concentration jump produced by photolysis of a protected derivative of ATP, caged ATP. K+ titrations were used to study the effect of K+ on the charge translocation kinetics of the protein. The time-dependent currents obtained after activation of the enzyme with caged ATP were analyzed with a simplified Albers-Post model (E1 (k1)-->E1ATP (k2)-->E2P (k3)-->E1) taking into account the capacitive coupling of the protein to the measuring system. The results of the K+ titrations show a strong dependence of the rate constant k3 on the K+ concentration at the extracellular side of the protein, indicating the K+ activated dephosphorylation reaction. In contrast, k1 and k2 remained constant. The K+ dependence of the rate k3 could be well described with a K+ binding model with two equivalent binding sites (E2P + 2K+ <==> E2P(K) + K+ <==> E2 P(2K)) followed by a rate limiting reaction (E2P(2K) --> E1(2K)). The half saturating K+ concentration K3,0.5 and the microscopic dissociation constant K3 for the K+ dependence of k3 were 4.5mM and 1.9mM respectively. At saturating K+ concentration the rate constant k3 was approximately 100 s(-1). The relative amount of net charge transported during the Na+ and the K+ dependent reactions could be determined from the experiments. Our results suggest electroneutral K+ translocation and do not support electrogenic K+ binding in an extracellular access channel. This is compatible with a model where 2 negative charges are cotransported with 3Na+ and 2K+ ions. Error analysis gives an upper limit of 20% charge transported during K+ translocation or during electrogenic K+ binding in a presumptive access channel compared to Na+ translocation.
    [Abstract] [Full Text] [Related] [New Search]