Interleukin-8 activates microtubule-associated protein 2 kinase (ERK1) in human neutrophils |
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Authors: | Van Lint Johan Van Damme Jo Billiau Alfons Merlevede Wilfried Vandenheede Jackie R |
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Institution: | (1) Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit te Leuven, Campus Gasthuisberg, B-3171, Belgium;(2) Rega Institute for Medical Research, Katholieke Universiteit te Leuven, Campus Gasthuisberg, B-3171, Belgium |
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Abstract: | The signal transduction initiated by the human cytokine interleukin-8 (IL-8), the main chemotactic cytokine for neutrophils, was investigated and found to encompass the stimulation of protein kinases. More specifically, IL-8 caused a transient, dose and time dependent activation of a Ser/Thr kinase activity towards myelin basic protein (MBP) and the MBP-derived peptide APRTPGGRR patterned after the specific concensus sequence in MBP for ERK enzymes. The activated MBP kinase was furthermore identified as an extracellular signal regulated kinase (ERK1) based on several criteria such as substrate specificity, molecular weight, activation-dependent mobility shift, and recognition by anti-ERK antibodies. For comparison, the chemotactic response of neutrophils to a stimulus of bacterial origin (fMet-Leu-Phe or fMLP) was also examined and found to involve the activation of a similar ERK enzyme. The present data clearly indicate that in terminally differentiated, non-proliferating human cells, the MBP kinase/ERK activity can serve other purposes than mitogenic signaling, and that processes such as chemotaxis, induced by bacterial peptides as well as by human cytokines like IL-8, involve the regulation of ERK enzyme.Abbreviations IL-8
interleukin-8
- fMLP
fMet-Leu-Phe
- MBP
myelin basic protein
- ERK
extracellular signal regulated kinase
- MAP2
microtubule-associated protein 2
- PK-A
cAMP dependent protein kinase
- PKI
protein kinase inhibitor
- PMSF
phenyl-methanesulfonyl fluoride
- PVDF
poly-vinylidene difluoride
- HBSF
Hank's buffered salt solution
- DAB
3,3-diaminobenzidine tetrahydrochloride
- PNPP
p-nitrophenyl-phosphate
- HSA
human serum albumin
- EGTA
ethylenebis (oxyethylenenitrilo)]tetraacetic acid
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis |
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Keywords: | interleukin-8 fMet-Leu-Phe human neutrophils chemotaxis MBP/MAP2 kinase/ERK1 signal transduction |
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