Reconstitution of recombinant N -formyl chemotactic peptide receptor with G protein

1993 ◽  
Vol 53 (4) ◽  
pp. 470-474 ◽  
Author(s):  
Ronda E. Schreiber ◽  
Eric R. Prossnitz ◽  
Richard D. Ye ◽  
Charles G. Cochrane ◽  
Algirdas J. Jesaitis ◽  
...  
Keyword(s):  
G Protein ◽  
Chemotactic Peptide ◽  
Peptide Receptor

scholarly journals Association of ligand-receptor complexes with actin filaments in human neutrophils: a possible regulatory role for a G-protein.

1989 ◽  
Vol 109 (6) ◽  
pp. 2791-2799 ◽  
Author(s):  
E Särndahl ◽  
M Lindroth ◽  
T Bengtsson ◽  
M Fällman ◽  
J Gustavsson ◽  
...  
Keyword(s):  
G Protein ◽  
Second Messengers ◽  
Human Neutrophils ◽  
Receptor Complex ◽  
Chemotactic Peptide ◽  
Filamentous Actin ◽  
High Concentration ◽  
Intact Cells ◽  
Peptide Receptor ◽  
Receptor Complexes

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.

Molecular Cloning and Chromosomal Localization of the MouseGpr37Gene Encoding an Orphan G-Protein-Coupled Peptide Receptor Expressed in Brain and Testis

Genomics ◽  
1998 ◽  
Vol 53 (3) ◽  
pp. 315-324 ◽  
Author(s):  
Daniela Marazziti ◽  
Angela Gallo ◽  
Elisabetta Golini ◽  
Rafaele Matteoni ◽  
Glauco P. Tocchini-Valentini,
Keyword(s):  
Molecular Cloning ◽  
G Protein ◽  
Chromosomal Localization ◽  
Peptide Receptor ◽  
G Protein Coupled

scholarly journals Characterization of an oligopeptide chemoattractant receptor on human blood monocytes using a new radioligand

Blood ◽  
1984 ◽  
Vol 63 (3) ◽  
pp. 588-592 ◽  
Author(s):  
MC Benyunes ◽  
R Snyderman
Keyword(s):  
Human Blood ◽  
Polymorphonuclear Leukocytes ◽  
Specific Radioactivity ◽  
Blood Monocytes ◽  
Chemotactic Peptide ◽  
Peripheral Blood Monocytes ◽  
Peptide Receptor ◽  
Human Polymorphonuclear Leukocytes ◽  
Dissociation Constant Kd

Abstract The study of chemoattractant receptors on human monocytes had been limited by the lack of a radioligand suitable for use with the small numbers of cells routinely available from human donors. A new synthetic oligopeptide radioligand f[35S]met-leu-phe, with a higher specific radioactivity than was available with the tritiated compound, was used to characterize a chemoattractant receptor on freshly isolated human blood monocytes. These cells bind f[35S]met-leu-phe with a dissociation constant (KD) of 30.2 +/- 5.6 nM and contain 84,000 +/- 11,300 receptors per cell. f[35S]met-leu-phe does not bind specifically to blood lymphocytes. The specificity of the oligopeptide receptor on monocytes is indistinguishable from the oligopeptide chemoattractant receptor on human polymorphonuclear leukocytes. Using f[35S]met-leu- phe, it will now be feasible to study the chemotactic peptide receptor on small numbers of partially purified peripheral blood monocytes from patients with defects of immune function.

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