Biochemical and Biophysical Research Communications
Identification of a free fatty acid receptor, FFA2R, expressed on leukocytes and activated by short-chain fatty acids
Section snippets
Materials and methods
Cloning. Cloning of the GPR43 ORF (open reading frame) from genomic (human and mouse) DNA was performed by PCR using the following primers:
Human ORF:
forward: 5′-ATTGCGGCCGCAGGATGCTGCCGGACTGGAAGAGC-3′;
reverse: 5′-ATTGCGGCCGCACTGCTACTCTGTAGTGAAGTCCGA-3′.
Murine ORF:
forward: 5′-CGGAATTCCGGGGGACTCTCTACTCGGTGACAA-3′;
reverse: 5′-ATTGCGGCCGCCAGAATGACCCCAGACTGGCACAGT-3′.
PCR conditions were: 96 °C for 3 min, 96 °C for 45 s; 58 °C for 1 min; and 72 °C for 1 min (30 cycles). The PCR products were restricted with
SCFA-induced activation of human GPR43
The previously identified putative GPCR ORF, GPR43 (GenBank Accession No. AF024690) [23] was cloned and stably expressed in the recombinant reporter cell line, HFF11, which is the second-generation reporter cell line [19]. A spectrum of SCFAs activated the receptor, but failed to induce any response in sham-transfected reporter cells. High concentrations, in the millimolar range, of SCFA, such as acetate (carbon chain length=C2), propionate (C3), and butyrate (C4), activated the reporter cells
Concluding remarks
What the actual physiological significance is and how SCFA activation of FFA2R affects the immune system, primarily the granulocytes, remain to be further investigated. SCFA have a multitude of cellular and immunoregulatory effects and, in fact, all related functions of PMNC, except cell adhesion, appear to be modulated by SCFA [25]. We have identified the previous orphan GPR43 as a receptor that is functionally activated by SCFA—most efficiently by acetate and propionate, but not by
Acknowledgements
This work was supported by GS Development, the Segerfalk Foundation, Crafoord Foundation, Ingabritt and Arne Lundberg Foundation, Kock Foundation, Swedish Society for Medical Research, Royal Physiographic Society, and Swedish Research Council.
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These authors contributed equally to this work.