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Association of ligand-receptor complexes with actin filaments in human neutrophils: a possible regulatory role for a G-protein.
- Source :
-
The Journal of cell biology [J Cell Biol] 1989 Dec; Vol. 109 (6 Pt 1), pp. 2791-9. - Publication Year :
- 1989
-
Abstract
- Most ligand-receptor interactions result in an immediate generation of various second messengers and a subsequent association of the ligand-receptor complex to the cytoskeleton. Depending on the receptor involved, this linkage to the cytoskeleton has been suggested to play a role in the termination of second messenger generation and/or the endocytic process whereby the ligand-receptor complex is internalized. We have studied how the binding of chemotactic peptide-receptor complexes to the cytoskeleton of human neutrophils is accomplished. As much as 76% of the tritiated formylmethionyl-leucyl-phenylalanine (fMet-Leu-[3H]Phe) specifically bound to intact cells, obtained by a 30-s stimulation with 20 nM fMet-Leu-[3H]Phe, still remained after Triton X-100 extraction. Preincubating intact cells with dihydrocytochalasin B (dhCB) or washing the cytoskeletal preparation with a high concentration of potassium, reduced the binding of ligand-receptor complexes to the cytoskeleton by 46% or more. Inhibition of fMet-Leu-Phe-induced generation of second messengers by ADP-ribosylating the alpha-subunit of the receptor-coupled G-protein with pertussis toxin, did not reduce the binding of ligand-receptor complexes to the cytoskeleton. However, using guanosine-5'-O-(2-thiodiphosphate) (GDP beta S) to prevent the dissociation of the fMet-Leu-Phe-associated G-protein within electrically permeabilized cells, led to a pronounced reduction (62%) of the binding between ligand-receptor complexes and the cytoskeleton. In summary, in human neutrophils the rapid association between chemotactic peptide-receptor complexes and the cytoskeleton is dependent on filamentous actin. This association is most likely regulated by the activation and dissociation of the fMet-Leu-Phe-associated G-protein.
- Subjects :
- Antibodies, Monoclonal
Cytochalasin B analogs & derivatives
Cytochalasin B pharmacology
Cytoskeleton drug effects
Cytoskeleton metabolism
Cytoskeleton ultrastructure
GTP-Binding Proteins physiology
Guanosine Diphosphate analogs & derivatives
Guanosine Diphosphate pharmacology
Humans
In Vitro Techniques
Kinetics
Ligands
Microscopy, Electron, Scanning
Neutrophils immunology
Pertussis Toxin
Potassium Chloride pharmacology
Receptors, Formyl Peptide
Receptors, Immunologic drug effects
Thionucleotides pharmacology
Virulence Factors, Bordetella pharmacology
Actins blood
GTP-Binding Proteins blood
N-Formylmethionine Leucyl-Phenylalanine metabolism
Neutrophils metabolism
Receptors, Immunologic metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9525
- Volume :
- 109
- Issue :
- 6 Pt 1
- Database :
- MEDLINE
- Journal :
- The Journal of cell biology
- Publication Type :
- Academic Journal
- Accession number :
- 2512299
- Full Text :
- https://doi.org/10.1083/jcb.109.6.2791