Refined Estimation of Kinetic Parameters of the Na+/H+ Antiport in Human Fibroblasts and Platelets

Jeffrey P. Gardner, Goro Tokudome, Haruo Tomonari, Mordechai Aladjem, Edward Cragoe, Abraham Aviv

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

A technique is presented to estimate the initial rates of Na+-dependent alkal-inization of acidified human fibroblasts and platelets and assess the kinetics of the Na+/ H+ antiport in these cells. Cytosolic pH (pHi) exhibits an exponential recovery following cellular acidification. Thus, the length of the time interval selected to monitor changes in pHi (ΔpHi) is critical to estimating the kinetics of the Na+/H+ antiport. We compared kinetic parameters of the Na+/H+ antiport, using computed and observed changes in ΔpHi, for arbitrarily selected time intervals following Na+-dependent activation. In both cells, significant increases in both the [Na+] for half-maximal activation (K 0.5) and maximal velocities (V max) were observed as ΔpHi was decreased. We conclude that kinetic parameters derived from initial rate determinations enable a more accurate characterization of the Na+/H+ antiport.

Original languageEnglish (US)
Pages (from-to)165-171
Number of pages7
JournalProceedings of the Society for Experimental Biology and Medicine
Volume194
Issue number2
DOIs
StatePublished - Jan 1 1990

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

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