Kinetic analysis of the binding of monomeric and dimeric ephrins to Eph receptors: Correlation to function in a growth cone collapse assay

Kumar B. Pabbisetty, Xin Yue, Chen Li, Juha Pekka Himanen, Renping Zhou, Dimitar B. Nikolov, Longqin Hu

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

Eph receptors and ephrins play important roles in regulating cell migration and positioning during both normal and oncogenic tissue development. Using a surface plasma resonance (SPR) biosensor, we examined the binding kinetics of representative monomeric and dimeric ephrins to their corresponding Eph receptors and correlated the apparent binding affinity with their functional activity in a neuronal growth cone collapse assay. Our results indicate that the Eph receptor binding of dimeric ephrins, formed through fusion with disulfide-linked Fc fragments, is best described using a bivalent analyte model as a two-step process involving an initial monovalent 2:1 binding followed by a second bivalent 2:2 binding. The bivalent binding dramatically decreases the apparent dissociation rate constants with little effect on the initial association rate constants, resulting in a 30-to 6000-fold decrease in apparent equilibrium dissociation constants for the binding of dimeric ephrins to Eph receptors relative to their monomeric counterparts. Interestingly, the change was more prominent in the A-class ephrin/Eph interactions than in the B-class of ephrins to Eph receptors. The increase in apparent binding affinities correlated well with increased activation of Eph receptors and the resulting growth cone collapse. Our kinetic analysis and correlation of binding affinity with function helped us better understand the interactions between ephrins and Eph receptors and should be useful in the design of inhibitors that interfere with the interactions. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish (US)
Pages (from-to)355-361
Number of pages7
JournalProtein Science
Volume16
Issue number3
DOIs
StatePublished - Mar 2007

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Keywords

  • Eph receptor
  • Ephrin
  • Growth cone collapse
  • Receptor dimerization
  • Surface plasmon resonance

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