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.
Harada
Masataka
M
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>
Habata
Yugo
Y
Hosoya
Masaki
M
Nishi
Kazunori
K
Fujii
Ryo
R
Kobayashi
Makoto
M
Hinuma
Shuji
S
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