PMID: 9169597May 1, 1997Paper

Kinetics of low-density lipoprotein receptor activity in Hep-G2 cells: derivation and validation of a Briggs-Haldane-based kinetic model for evaluating receptor-mediated endocytotic processes in which receptors recycle

The Biochemical Journal
H J Harwood, L D Pellarin

Abstract

The process of receptor-mediated endocytosis for receptors that recycle to the cell surface in an active form can be considered as being kinetically analogous to that of a uni-substrate, uni-product enzyme-catalysed reaction. In this study we have derived steady-state initial-velocity rate equations for this process, based on classical Briggs-Haldane and King-Altman kinetic approaches to multi-step reactions, and have evaluated this kinetic paradigm, using as a model system the low-density lipoprotein (LDL)-receptor-mediated endocytosis of the trapped label [14C]sucrose-LDL in uninduced, steady-state Hep-G2 cells. Using the derived rate equations, together with experimentally determined values for Bmax (123 fmol/mg of cell protein), Kd (14.3 nM), the endocytotic rate constant ke (analogous to kcat; 0.163 min-1), Km (80 nM) and maximal internalization velocity (26.4 fmol/min per mg), we have calculated the ratio ke/Km (0.00204 nM-1.min-1), the bimolecular rate constant for LDL and LDL-receptor association (0. 00248 nM-1.min-1), the first-order rate constant for LDL-LDL-receptor complex dissociation (0.0354 min-1), the total cellular content of LDL receptors (154 fmol/mg of cell protein), the intracellular LDL receptor concentrat...Continue Reading

Citations

Jul 15, 2009·Molecular Oncology·Marc R Birtwistle, Boris N Kholodenko
Feb 22, 2002·European Journal of Immunology·Charlotte MennéCarsten Geisler
May 12, 2004·Biotechnology and Bioengineering·Kwang-Il LimJohn Yin
Aug 22, 2008·Bulletin of Mathematical Biology·J A D WattisK G Jackson
Apr 27, 2017·Journal of Lipid Research·Seyedeh Maryam ZekavatNorman A Mazer
Aug 16, 2008·Journal of Mathematical Biology·T PearsonH M Byrne
Sep 26, 2017·PLoS Computational Biology·Stefan M KallenbergerDirk-Peter Herten
Jan 18, 2003·Journal of Pharmaceutical Sciences·M SarntinoranontP F Morrison
Jun 29, 2004·Journal of Biotechnology·Noppol LeksawasdiPeter L Rogers
May 15, 2015·Bioconjugate Chemistry·Tang TangAngelique Y Louie
Dec 27, 2008·Journal of Theoretical Biology·M J TindallK G Jackson
Mar 16, 2006·Bioconjugate Chemistry·Björn GustafssonAngelique Y Louie

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