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Regulation of human interleukin-8 receptor A: identification of a phosphorylation site involved in modulating receptor functions.

Publication ,  Journal Article
Richardson, RM; DuBose, RA; Ali, H; Tomhave, ED; Haribabu, B; Snyderman, R
Published in: Biochemistry
October 31, 1995

The human type A interleukin-8 receptor (IL-8RA) was modified to express an amino-terminal epitope tag and stably overexpressed in a rat basophilic leukemia cell line (RBL-2H3). This receptor (ET-IL-8RA) displayed functional properties similar to those of the native receptor in neutrophils in that exposure to IL-8 stimulated GTPase activity, phosphoinositide (PI) hydrolysis, intracellular calcium mobilization, and degranulation in a pertussis toxin (PTx) susceptible fashion. IL-8 induced dose- and time-dependent phosphorylation of ET-IL-8RA. Phorbol 12-myristate 13-acetate (PMA) treatment also resulted in phosphorylation of the receptor although to a lesser extent. Staurosporine totally blocked PMA-induced phosphorylation but only partially inhibited IL-8-mediated phosphorylation. Phosphorylation of ET-IL-8RA correlated with its desensitization as measured by GTPase activation and calcium mobilization. To determine the role of phosphorylation in IL-8RA signal transduction, three mutants lacking specific serine and threonine residues located at the C-terminal of this receptor were constructed by site-directed mutagenesis (M1, M2, and M3). The mutated receptors expressed in RBL-2H3 cells displayed pharmacological properties (Kd approximately 2-2.8 nM and Bmax approximately 3-3.5 pmol/mg of protein) similar to those of the wild-type ET-IL-8RA. M2 and M3, but not M1, showed a marked decrease in IL-8-induced phosphorylation compared to the wild-type receptor. M2 and M3 but not M1 were resistant to PMA-mediated phosphorylation and desensitization and were also more resistant to homologous desensitization than M1 or ET-IL-8RA.(ABSTRACT TRUNCATED AT 250 WORDS)

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Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

October 31, 1995

Volume

34

Issue

43

Start / End Page

14193 / 14201

Location

United States

Related Subject Headings

  • Up-Regulation
  • Tumor Cells, Cultured
  • Threonine
  • Tetradecanoylphorbol Acetate
  • Staurosporine
  • Serine
  • Receptors, Interleukin-8A
  • Receptors, Interleukin
  • Rats
  • Protein Kinase C
 

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Richardson, R. M., DuBose, R. A., Ali, H., Tomhave, E. D., Haribabu, B., & Snyderman, R. (1995). Regulation of human interleukin-8 receptor A: identification of a phosphorylation site involved in modulating receptor functions. Biochemistry, 34(43), 14193–14201. https://doi.org/10.1021/bi00043a025
Richardson, R. M., R. A. DuBose, H. Ali, E. D. Tomhave, B. Haribabu, and R. Snyderman. “Regulation of human interleukin-8 receptor A: identification of a phosphorylation site involved in modulating receptor functions.Biochemistry 34, no. 43 (October 31, 1995): 14193–201. https://doi.org/10.1021/bi00043a025.
Richardson RM, DuBose RA, Ali H, Tomhave ED, Haribabu B, Snyderman R. Regulation of human interleukin-8 receptor A: identification of a phosphorylation site involved in modulating receptor functions. Biochemistry. 1995 Oct 31;34(43):14193–201.
Richardson, R. M., et al. “Regulation of human interleukin-8 receptor A: identification of a phosphorylation site involved in modulating receptor functions.Biochemistry, vol. 34, no. 43, Oct. 1995, pp. 14193–201. Pubmed, doi:10.1021/bi00043a025.
Richardson RM, DuBose RA, Ali H, Tomhave ED, Haribabu B, Snyderman R. Regulation of human interleukin-8 receptor A: identification of a phosphorylation site involved in modulating receptor functions. Biochemistry. 1995 Oct 31;34(43):14193–14201.
Journal cover image

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

October 31, 1995

Volume

34

Issue

43

Start / End Page

14193 / 14201

Location

United States

Related Subject Headings

  • Up-Regulation
  • Tumor Cells, Cultured
  • Threonine
  • Tetradecanoylphorbol Acetate
  • Staurosporine
  • Serine
  • Receptors, Interleukin-8A
  • Receptors, Interleukin
  • Rats
  • Protein Kinase C