Subcellular Localization and Translocation of the Receptor for N-formylmethionyl-leucyl-phenylalanine in Human Neutrophils

Overview

Journal Biochem J

Specialty Biochemistry

Date 1994 Apr 15

PMID 8172608

Citations 37

Authors

H Sengelov

F Boulay

L Kjeldsen

N Borregaard

Affiliations

Soon will be listed here.

Abstract

The subcellular localization of N-formylmethionyl-leucyl-phenylalanine (fMLP) receptors in human neutrophils was investigated. The fMLP receptor was detected with a high-affinity, photoactivatable, radioiodinated derivative of N-formyl-methionyl-leucyl-phenylalanyl-lysine (fMLFK). Neutrophils were disrupted by nitrogen cavitation and fractionated on Percoll density gradients. fMLP receptors were located in the beta-band containing gelatinase and specific granules, and in the gamma-band containing plasma membrane and secretory vesicles. Plasma membranes and secretory vesicles were separated by high-voltage free-flow electrophoresis, and secretory vesicles were demonstrated to be highly enriched in fMLP receptors. The receptors found in secretory vesicles translocated fully to the plasma membrane upon stimulation with inflammatory mediators. The receptor translocation from the beta-band indicated that the receptor present there was mainly located in gelatinase granules. A 25 kDa fMLP-binding protein was found in the beta-band. Immunoprecipitation revealed that this protein was identical with NGAL (neutrophil gelatinase-associated lipocalin), a novel protein found in specific granules. In summary, we demonstrate that the compartment in human neutrophils that is mobilized most easily and fastest, the secretory vesicle, is a major reservoir of fMLP receptors. This explains the prompt and extensive upregulation of fMLP receptors on the neutrophil surface in response to inflammatory stimuli.

Citing Articles

Neutrophil diversity and function in health and disease.

Zhang F, Xia Y, Su J, Quan F, Zhou H, Li Q Signal Transduct Target Ther. 2024; 9(1):343.

PMID: 39638788 PMC: 11627463. DOI: 10.1038/s41392-024-02049-y.

Increased expression of formyl peptide receptor-1 by basophils from patients with mastocytosis.

Yin Y, Li J, Metcalfe D, Olivera A, Frischmeyer-Guerrerio P, Carter M J Allergy Clin Immunol Glob. 2024; 3(4):100296.

PMID: 39148513 PMC: 11325225. DOI: 10.1016/j.jacig.2024.100296.

ATG5 provides host protection acting as a switch in the atg8ylation cascade between autophagy and secretion.

Wang F, Peters R, Jia J, Mudd M, Salemi M, Allers L Dev Cell. 2023; 58(10):866-884.e8.

PMID: 37054706 PMC: 10205698. DOI: 10.1016/j.devcel.2023.03.014.

Therapeutic assessment of N-formyl-methionyl-leucyl-phenylalanine (fMLP) in reducing periprosthetic joint infection.

Hamilton J, Mohamed M, Witt B, Wimmer M, Shafikhani S Eur Cell Mater. 2021; 42:122-138.

PMID: 34435345 PMC: 8459619. DOI: 10.22203/eCM.v042a09.

Mitochondrial N-formyl methionine peptides associate with disease activity as well as contribute to neutrophil activation in patients with rheumatoid arthritis.

Duvvuri B, Baddour A, Deane K, Feser M, Nelson J, Demoruelle M J Autoimmun. 2021; 119:102630.

PMID: 33713887 PMC: 8062236. DOI: 10.1016/j.jaut.2021.102630.

References

Boyum A . Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968; 97:77-89. View

Graves V, Gabig T, McCarthy L, Strour E, Leemhuis T, English D . Simultaneous mobilization of Mac-1 (CD11b/CD18) and formyl peptide chemoattractant receptors in human neutrophils. Blood. 1992; 80(3):776-87. View

Dechatelet L, Cooper M . A modified procedure for the determination of leukocyte alkaline phosphatase. Biochem Med. 1970; 4(1):61-8. DOI: 10.1016/0006-2944(70)90103-1. View

Schiffmann E, CORCORAN B, Wahl S . N-formylmethionyl peptides as chemoattractants for leucocytes. Proc Natl Acad Sci U S A. 1975; 72(3):1059-62. PMC: 432465. DOI: 10.1073/pnas.72.3.1059. View

Gould R, Ginsberg B, Spector A . Effects of octyl beta-glucoside on insulin binding to solubilized membrane receptors. Biochemistry. 1981; 20(24):6776-81. DOI: 10.1021/bi00527a006. View

Jesaitis A, Naemura J, Painter R, Sklar L, Cochrane C . Intracellular localization of N-formyl chemotactic receptor and Mg2+ dependent ATPase in human granulocytes. Biochim Biophys Acta. 1982; 719(3):556-68. DOI: 10.1016/0304-4165(82)90246-x. View

Borregaard N, Heiple J, Simons E, Clark R . Subcellular localization of the b-cytochrome component of the human neutrophil microbicidal oxidase: translocation during activation. J Cell Biol. 1983; 97(1):52-61. PMC: 2112494. DOI: 10.1083/jcb.97.1.52. View

Fletcher M, Gallin J . Human neutrophils contain an intracellular pool of putative receptors for the chemoattractant N-formyl-methionyl-leucyl-phenylalanine. Blood. 1983; 62(4):792-9. View

Andersson T, Dahlgren C, Lew P, Stendahl O . Cell surface expression of fMet-Leu-Phe receptors on human neutrophils. Correlation to changes in the cytosolic free Ca2+ level and action of phorbol myristate acetate. J Clin Invest. 1987; 79(4):1226-33. PMC: 424318. DOI: 10.1172/JCI112941. View

10.

Perez H, Elfman F, Lobo E . Removal of human polymorphonuclear leukocyte surface sialic acid inhibits reexpression (or recycling) of formyl peptide receptors. A possible explanation for its effect on formyl peptide-induced polymorphonuclear leukocyte chemotaxis. J Immunol. 1987; 139(6):1978-84. View

11.

Borregaard N, Miller L, Springer T . Chemoattractant-regulated mobilization of a novel intracellular compartment in human neutrophils. Science. 1987; 237(4819):1204-6. DOI: 10.1126/science.3629236. View

12.

Rotrosen D, Gallin J, Spiegel A, Malech H . Subcellular localization of Gi alpha in human neutrophils. J Biol Chem. 1988; 263(22):10958-64. View

13.

English D, Broxmeyer H, Gabig T, Akard L, Williams D, Hoffman R . Temporal adaptation of neutrophil oxidative responsiveness to n-formyl-methionyl-leucyl-phenylalanine. Acceleration by granulocyte-macrophage colony stimulating factor. J Immunol. 1988; 141(7):2400-6. View

14.

Tennenberg S, Zemlan F, Solomkin J . Characterization of N-formyl-methionyl-leucyl-phenylalanine receptors on human neutrophils. Effects of isolation and temperature on receptor expression and functional activity. J Immunol. 1988; 141(11):3937-44. View

15.

Volpp B, Nauseef W, Clark R . Subcellular distribution and membrane association of human neutrophil substrates for ADP-ribosylation by pertussis toxin and cholera toxin. J Immunol. 1989; 142(9):3206-12. View

16.

Allen R, Erickson R, Jesaitis A . Identification of a human neutrophil protein of Mr 24 000 that binds N-formyl peptides: co-sedimentation with specific granules. Biochim Biophys Acta. 1989; 991(1):123-33. DOI: 10.1016/0304-4165(89)90037-8. View

17.

Bjerrum O, Borregaard N . Dual granule localization of the dormant NADPH oxidase and cytochrome b559 in human neutrophils. Eur J Haematol. 1989; 43(1):67-77. DOI: 10.1111/j.1600-0609.1989.tb01254.x. View

18.

Kjeldsen L, Bjerrum O, Askaa J, Borregaard N . Subcellular localization and release of human neutrophil gelatinase, confirming the existence of separate gelatinase-containing granules. Biochem J. 1992; 287 ( Pt 2):603-10. PMC: 1133208. DOI: 10.1042/bj2870603. View

19.

Kjeldsen L, Bjerrum O, Hovgaard D, Johnsen A, Sehested M, Borregaard N . Human neutrophil gelatinase: a marker for circulating blood neutrophils. Purification and quantitation by enzyme linked immunosorbent assay. Eur J Haematol. 1992; 49(4):180-91. DOI: 10.1111/j.1600-0609.1992.tb00045.x. View

20.

Sengelov H, Kjeldsen L, Borregaard N . Control of exocytosis in early neutrophil activation. J Immunol. 1993; 150(4):1535-43. View