title: Kinetics of leptin binding to the Q223R leptin receptor.
abstract: Studies in human populations and mouse models of disease have linked the common leptin receptor Q223R mutation to obesity, multiple forms of cancer, adverse drug reactions, and susceptibility to enteric and respiratory infections. Contradictory results cast doubt on the phenotypic consequences of this variant. We set out to determine whether the Q223R substitution affects leptin binding kinetics using surface plasmon resonance (SPR), a technique that allows sensitive real-time monitoring of protein-protein interactions. We measured the binding and dissociation rate constants for leptin to the extracellular domain of WT and Q223R murine leptin receptors expressed as Fc-fusion proteins and found that the mutant receptor does not significantly differ in kinetics of leptin binding from the WT leptin receptor. (WT: ka 1.76×106±0.193×106 M-1 s-1, kd 1.21×10-4±0.707×10-4 s-1, KD 6.47×10-11±3.30×10-11 M; Q223R: ka 1.75×106±0.0245×106 M-1 s-1, kd 1.47×10-4±0.0505×10-4 s-1, KD 8.43×10-11±0.407×10-11 M). Our results support earlier findings that differences in affinity and kinetics of leptin binding are unlikely to explain mechanistically the phenotypes that have been linked to this common genetic variant. Future studies will seek to elucidate the mechanism by which this mutation influences susceptibility to metabolic, infectious, and malignant pathologies.
authors.affiliation: Department of Medicine, Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, United States of America.
authors.forename: Hans
authors.initials: H
authors.lastname: Verkerke
authors.affiliation: Department of Medicine, Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, United States of America.
authors.forename: Caitlin
authors.initials: C
authors.lastname: Naylor
authors.affiliation: Flanders Institute for Biotechnology (VIB), Department of Medical Protein Research, Ghent University, Ghent, Belgium.
authors.forename: Lennart
authors.initials: L
authors.lastname: Zabeau
authors.affiliation: Flanders Institute for Biotechnology (VIB), Department of Medical Protein Research, Ghent University, Ghent, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Tavernier
authors.affiliation: Department of Medicine, Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, United States of America.
authors.forename: William A
authors.initials: WA
authors.lastname: Petri
authors.affiliation: Department of Medicine, Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, United States of America.
authors.forename: Chelsea
authors.initials: C
authors.lastname: Marie
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D020738:Leptin / Q000378:metabolism*; D051379:Mice; D050505:Mutant Proteins / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding; D054411:Receptors, Leptin / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020349:Surface Plasmon Resonance
chemical_list: D020738:Leptin; D050505:Mutant Proteins; D054411:Receptors, Leptin; D011993:Recombinant Fusion Proteins
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