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Mol. Cells 2007; 24(1): 119-124
Published online January 1, 1970
© The Korean Society for Molecular and Cellular Biology
Kinetics of Binding of LPS to Recombinant CD14, TLR4, and MD-2 Proteins
TLR4 together with CD14 and MD-2 forms a pattern recognition receptor that plays an initiating role in the innate immune response to Gram-negative bacteria. Here, we employed the surface plasmon resonance technique to investigate the kinetics of binding of LPS to recombinant CD14, MD-2 and TLR4 proteins produced in insect cells. The dissociation constants (KD) of LPS for immobilized CD14 and MD-2 were 8.7 ?M, and 2.3 ?M, respectively. The association rate constant (Kon) of LPS for MD-2 was 5.61 ? 103 M?1S?1, and the dissociation rate constant (Koff) was 1.28 ? 10?2 S?1, revealing slow association and fast dissociation with an affinity constant KD of 2.33 ? 10?6 M at 25?C. These affinities are consistent with the current view that CD14 conveys LPS to the TLR4/MD-2 complex.
Keywords Affinity; CD14; LPS; MD-2; Surface Plasmon Resonance; TLR4.
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Research Article
Mol. Cells 2007; 24(1): 119-124
Published online August 31, 2007
Copyright © The Korean Society for Molecular and Cellular Biology.
Kinetics of Binding of LPS to Recombinant CD14, TLR4, and MD-2 Proteins
Han Jae Shin, Hayyoung Lee, Jong Dae Park, Hak Chul Hyun, Hyung Ok Sohn, Dong Wook Lee and Young Sang Kim
Abstract
TLR4 together with CD14 and MD-2 forms a pattern recognition receptor that plays an initiating role in the innate immune response to Gram-negative bacteria. Here, we employed the surface plasmon resonance technique to investigate the kinetics of binding of LPS to recombinant CD14, MD-2 and TLR4 proteins produced in insect cells. The dissociation constants (KD) of LPS for immobilized CD14 and MD-2 were 8.7 ?M, and 2.3 ?M, respectively. The association rate constant (Kon) of LPS for MD-2 was 5.61 ? 103 M?1S?1, and the dissociation rate constant (Koff) was 1.28 ? 10?2 S?1, revealing slow association and fast dissociation with an affinity constant KD of 2.33 ? 10?6 M at 25?C. These affinities are consistent with the current view that CD14 conveys LPS to the TLR4/MD-2 complex.
Keywords: Affinity, CD14, LPS, MD-2, Surface Plasmon Resonance, TLR4.
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