Formyl peptide receptor like 1 differentially requires mitogen-activated protein kinases for the induction of glial fibrillary acidic protein and interleukin-1α in human U87 astrocytoma cells

2007 ◽  
Vol 19 (10) ◽  
pp. 2106-2117 ◽  
Author(s):  
Angel Y.F. Kam ◽  
Timothy T.M. Tse ◽  
Dawna H.T. Kwan ◽  
Yung H. Wong
Keyword(s):  
Glial Fibrillary Acidic Protein ◽  
Protein Kinases ◽  
Acidic Protein ◽  
Formyl Peptide Receptor ◽  
Mitogen Activated Protein Kinases ◽  
Peptide Receptor ◽  
Astrocytoma Cells ◽  
Formyl Peptide ◽  
Mitogen Activated Protein ◽  
Interleukin 1Α

scholarly journals A single amino acid substitution (N297A) in the conserved NPXXY sequence of the human N-formyl peptide receptor results in inhibition of desensitization and endocytosis, and a dose-dependent shift in p42/44 mitogen-activated protein kinase activation and chemotaxis

2000 ◽  
Vol 352 (2) ◽  
pp. 399-407 ◽  
Author(s):  
Jeannie M. GRIPENTROG ◽  
Algirdas J. JESAITIS ◽  
Heini M. MIETTINEN
Keyword(s):  
Protein Kinase ◽  
Transmembrane Domain ◽  
Mitogen Activated Protein Kinase ◽  
Formyl Peptide Receptor ◽  
Wild Type ◽  
Peptide Receptor ◽  
Formyl Peptide ◽  
Mitogen Activated Protein ◽  
Dose Dependent Increase ◽  
Dose Dependent

The formyl peptide receptor (FPR) is a G-protein-coupled receptor (GPCR) that mediates chemotaxis and stimulates the mitogen-activated protein kinase (MAPK)/extracellular-signal-regulated kinase pathway. We have examined the functional effects of substitutions of a conserved aspartic acid residue in the second transmembrane domain (D71A) and of residues in the conserved NPXXY motif in the seventh transmembrane domain (N297A and Y301A). These mutated receptors, expressed in Chinese hamster ovary (CHO) cells, bind ligand with affinities similar to wild-type FPR, but the D71A mutant is uncoupled from G-protein [Miettinen, Mills, Gripentrog, Dratz, Granger and Jesaitis (1997) J. Immunol 159, 4045–4054]. In the present study, we show that both the D71A and N297A mutations resulted in defective endocytosis. The N297A substitution also prevented desensitization, as determined by intracellular calcium mobilization by sequential stimulation with ligand. In chemotaxis assays, the N297A mutation resulted in cell migration towards gradients of up to 100nM N-formyl-methionyl-leucyl-phenylalanine (fMLF), whereas cells expressing the wild-type FPR and the Y301A mutant were no longer chemotactically responsive at 10–100nM fMLF. Maximal activation of p42/44 MAPK occurred in CHO cells expressing wild-type FPR at 10nM–100nM fMLF, whereas cells expressing the N297A mutant showed a dose-dependent increase in the amount of phosphorylated p42/44 MAPK up to 1–10µM fMLF. Since the MAPK kinase inhibitor PD98059 blocked fMLF-induced chemotaxis, our results suggest that the dose-dependent increase in p42/44 MAPK activation may correlate with the increased chemotactic migration of N297A transfectants at 10nM–100nM fMLF.

Activation of multiple mitogen-activated protein kinases by recombinant calcitonin gene-related peptide receptor

2000 ◽  
Vol 389 (2-3) ◽  
pp. 125-130 ◽  
Author(s):  
Narayanan Parameswaran ◽  
Jyoti Disa ◽  
William S. Spielman ◽  
David P. Brooks ◽  
Ponnal Nambi ◽  
...  
Keyword(s):  
Protein Kinases ◽  
Calcitonin Gene Related Peptide ◽  
Mitogen Activated Protein Kinases ◽  
Peptide Receptor ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Mitogen Activated Protein

scholarly journals EphrinBs/EphBs Signaling Is Involved in Modulation of Spinal Nociceptive Processing through a Mitogen-activated Protein Kinases-dependent Mechanism

2010 ◽  
Vol 112 (5) ◽  
pp. 1234-1249 ◽  
Author(s):  
Jia-Ping Ruan ◽  
Hong-Xing Zhang ◽  
Xian-Fu Lu ◽  
Yue-Peng Liu ◽  
Jun-Li Cao
Keyword(s):  
Glial Fibrillary Acidic Protein ◽  
Protein Kinases ◽  
Intrathecal Injection ◽  
Immunofluorescence Staining ◽  
Mitogen Activated Protein Kinases ◽  
Pain Behaviors ◽  
Neuronal Nuclei ◽  
Mitogen Activated Protein ◽  
Nociceptive Information ◽  
Fos Expression

Background Our previous studies have demonstrated that EphBs receptors and ephrinBs ligands were involved in modulation of spinal nociceptive information. However, the downstream mechanisms that control this process are not well understood. The aim of this study was to further investigate whether mitogen-activated protein kinases (MAPKs), as the downstream effectors, participate in modulation of spinal nociceptive information related to ephrinBs/EphBs. Methods Thermal hyperalgesia and mechanical allodynia were measured using radiant heat and von Frey filaments test. Immunofluorescence staining was used to detect the expression of p-MAPKs and of p-MAPKs/neuronal nuclei, or p-MAPKs/glial fibrillary acidic protein double label. C-Fos expression was determined by immunohistochemistry. The expression of p-MAPKs was also determined by Western blot assay. Results Intrathecal injection of ephrinB1-Fc produced a dose- and time-dependent thermal and mechanical hyperalgesia, accompanied by the increase of spinal p-MAPKs and c-Fos expression. Immunofluorescence staining revealed that p-MAPKs colocalized with the neuronal marker (neuronal nuclei) and the astrocyte marker (glial fibrillary acidic protein). Inhibition of MAPKs prevented and reversed pain behaviors and the increase of spinal c-Fos expression induced by intrathecal injection of ephrinB1-Fc. Inhibition of EphBs receptors by intrathecal injection of EphB1-Fc reduced formalin-induced inflammation and chronic constrictive injury-induced neuropathic pain behaviors accompanied by decreased expression of spinal p-MAPKs and c-Fos protein. Furthermore, pretreatment with MK-801, an N-methyl-d-aspartate receptor antagonist, prevented behavioral hyperalgesia and activation of spinal MAPKs induced by intrathecal injection of ephrinB1-Fc. Conclusions These results demonstrated that activation of MAPKs contributed to modulation of spinal nociceptive information related to ephrinBs/EphBs.

scholarly journals V101L of human formyl peptide receptor 1 (FPR1) increases receptor affinity and augments the antagonism mediated by cyclosporins

2013 ◽  
Vol 451 (2) ◽  
pp. 245-255 ◽  
Author(s):  
Caihong Zhou ◽  
Yan Zhou ◽  
Jia Wang ◽  
Yang Feng ◽  
Haonan Wang ◽  
...  
Keyword(s):  
Immunosuppressive Agent ◽  
Han Chinese ◽  
Mitogen Activated Protein Kinase ◽  
Formyl Peptide Receptor ◽  
Single Amino Acid ◽  
Nucleotide Polymorphisms ◽  
Receptor Affinity ◽  
Peptide Receptor ◽  
Formyl Peptide ◽  
Mitogen Activated Protein

Genetic variation plays a major role in drug response variability. CsA (cyclosporin A), a widely used immunosuppressive agent, is a specific antagonist for FPR1 (formyl peptide receptor 1), which is an important G-protein-coupled chemoattractant receptor in the innate immune system. In order to study the variable responses of cyclosporins to different FPR1 mutants, we investigated the distribution of human FPR1 haplotypes among 209 healthy Han Chinese subjects. The haplotype pattern in Han Chinese were characterized on the basis of five SNPs (single nucleotide polymorphisms), including rs5030878 (p.T11I), rs2070745 (p.V101L), rs5030880 (p.R190W), rs1042229 (p.N192K) and rs867228 (p.A346E). Receptor binding affinity of cyclosporins to FPR1 haplotypes was assessed using N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys–FITC in CHO-Gα16 cells stably transfected with cDNAs encoding the top 12 FPR1 haplotypes in the Han Chinese. Variants of FPR1 carrying a single amino acid substitution of leucine for valine at position 101 (p.Leu101) displayed significantly higher pKi values for CsA and CsH (cyclosporin H), indicative of an improved receptor affinity. The polymorphism of FPR1 p.Leu101 also enhanced the inhibitory effects of cyclosporins on fMLF (N-formyl-methionyl-leucyl-phenylalanine)-induced activities, including calcium mobilization, cell chemotaxis and MAPK (mitogen-activated protein kinase) phosphorylation. These results point to a possible complication for clinical use of CsA in patients carrying the p.Leu101 allele of FPR1.

Sign in / Sign up

Export Citation Format

Share Document