Semisynthesis and Application of Carboxyfluorescein-Labelled Biologically Active Human Interleukin-8
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R. David
Abstract
Human interleukin 8 (hIL-8), a neutrophil-activating and chemotactic cytokine, is known to play an important role in the pathogenesis of a large number of neutrophil-driven inflammatory diseases. This cytokine belongs to the family of CXC chemokines, mediating the response through binding to the seventransmembrane helical G protein-coupled receptors CXCR1 and CXCR2. For the first time, we employed the expressed protein ligation (EPL) strategy to chemokine synthesis and subsequent modification. The ligation site was chosen with respect to the position of four cysteine residues within the hIL-8 sequence. Ligation with synthetic peptides that carry cysteine at their N-termini resulted in fulllength hIL-8 and the specifically carboxyfluoresceinlabelled analogue [K69(CF)]hIL-8(1 77). [K69(CF)]hIL-8(1-77) was fully active as shown by inhibition of cAMP production. Furthermore, this analogue was used to study receptor internalisation in human promyelotic HL60 cells that express CXCR1 and CXCR2 receptors. Binding and quenching studies were performed on HL60 membranes and suggest that the Cterminus of IL-8 is accessible to solvent in the receptor-bound state. Thus, we introduce here a powerful approach that allows the site-specific incorporation of chemical modifications into the sequence of chemokines, which opens new avenues for studying IL-8 function.
Copyright © 2003 by Walter de Gruyter GmbH & Co. KG
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- Anti-Mouse GPI-PLD Antisera Highlight Structural Differences between Murine and Bovine GPI-PLDs
- Substrate Specificity of Glutaminyl Cyclases from Plants and Animals
- The Binding of Haptoglobin to Apolipoprotein AI: Influence of Hemoglobin and Concanavalin A
- A Novel Water-Soluble and Cell-Permeable Calpain Inhibitor Protects Myocardial and Mitochondrial Function in Postischemic Reperfusion
- Potent Bivalent Inhibition of Human Tryptase-? by a Synthetic Inhibitor
- Aza-Peptide Epoxides: Potent and Selective Inhibitors of Schistosoma mansoni and Pig Kidney Legumains (Asparaginyl Endopeptidases)
- Semisynthesis and Application of Carboxyfluorescein-Labelled Biologically Active Human Interleukin-8
- Acknowledgment
- Content Index
- Author Index
- Subject Index
Articles in the same Issue
- Metabolic Stability, Receptor Binding, cAMP Generation, Insulin Secretion and Antihyperglycaemic Activity of Novel N-Terminal Glu9-Substituted Analogues of Glucagon-Like Peptide-1
- Characterisation of Human Dipeptidyl Peptidase IV Expressed in Pichia pastoris. A Structural and Mechanistic Comparison between the Recombinant Human and the Purified Porcine Enzyme
- Effects of Iron Limitation on the Respiratory Chain and the Membrane Cytochrome Pattern of the Euryarchaeon Halobacterium salinarum
- Anti-Mouse GPI-PLD Antisera Highlight Structural Differences between Murine and Bovine GPI-PLDs
- Substrate Specificity of Glutaminyl Cyclases from Plants and Animals
- The Binding of Haptoglobin to Apolipoprotein AI: Influence of Hemoglobin and Concanavalin A
- A Novel Water-Soluble and Cell-Permeable Calpain Inhibitor Protects Myocardial and Mitochondrial Function in Postischemic Reperfusion
- Potent Bivalent Inhibition of Human Tryptase-? by a Synthetic Inhibitor
- Aza-Peptide Epoxides: Potent and Selective Inhibitors of Schistosoma mansoni and Pig Kidney Legumains (Asparaginyl Endopeptidases)
- Semisynthesis and Application of Carboxyfluorescein-Labelled Biologically Active Human Interleukin-8
- Acknowledgment
- Content Index
- Author Index
- Subject Index
