N-Formylated humanin activates both formyl peptide receptor-like 1 and 2.
We have discovered that humanin (HN) acts as a ligand for formyl peptide receptor-like 1 (FPRL1) and 2 (FPRL2). This discovery was based on our finding that HN suppressed forskolin-induced cAMP production in Chinese hamster ovary (CHO) cells expressing human FPRL1 (CHO-hFPRL1) or human FPRL2 (CHO-hFPRL2). In addition, we found that N-formylated HN (fHN) performed more potently as a ligand for FPRL1 than HN: in CHO-hFPRL1 cells, the effective concentration for the half-maximal response (EC(50)) value of HN was 3.5nM, while that of fHN was 0.012nM. We demonstrated by binding experiments using [(125)I]-W peptide that HN and fHN directly interacted with hFPRL1 on the membrane. In addition, we found that HN and fHN showed strong chemotactic activity for CHO-hFPRL1 and CHO-hFPRL2 cells. HN is known to have a protective effect against neuronal cell death. Our findings contribute to the understanding of the mechanism behind HN's function.
Discovery Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Ltd., 10 Wadai, Tsukuba, Ibaraki 300-4293, Japan. Harada_Masataka@takeda.co.jp <Harada_Masataka@takeda.co.jp>
Masataka
M
Harada
Yugo
Y
Habata
Masaki
M
Hosoya
Kazunori
K
Nishi
Ryo
R
Fujii
Makoto
M
Kobayashi
Shuji
S
Hinuma
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D010446:Peptide Fragments / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D044042:Receptors, Formyl Peptide / Q000235:genetics / Q000378:metabolism*; D044067:Receptors, Lipoxin / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution
C105101:FPR2 protein, human; C469985:FPR3 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D010446:Peptide Fragments; D011506:Proteins; D044042:Receptors, Formyl Peptide; D044067:Receptors, Lipoxin; D011993:Recombinant Fusion Proteins; C427211:humanin; D000242:Cyclic AMP