RT Journal Article
SR Electronic
T1 The PAR2 signal peptide prevents premature receptor cleavage and activation
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 760769
DO 10.1101/760769
A1 Belinda Liu
A1 Grace Lee
A1 Jiejun Wu
A1 Janise Deming
A1 Chester Kuei
A1 Anthony Harrington
A1 Lien Wang
A1 Jennifer Towne
A1 Timothy Lovenberg
A1 Changlu Liu
A1 Siquan Sun
YR 2019
UL http://biorxiv.org/content/early/2019/09/06/760769.abstract
AB Unlike closely related GPCRs, protease-activated receptors (PAR1, PAR2, PAR3, and PAR4) have a predicted signal peptide at their N-terminus, which is encoded by a separate exon, suggesting that the signal peptides of PARs may serve an important and unique function, specific for PARs. In this report, we show that the PAR2 signal peptide, when fused to the N-terminus of IgG-Fc, effectively induced IgG-Fc secretion into culture medium, thus behaving like a classical signal peptide. The presence of PAR2 signal peptide has a strong effect on PAR2 cell surface expression, as deletion of the signal peptide (PAR2ΔSP) led to dramatic reduction of the cell surface expression and decreased responses to trypsin or the synthetic peptide ligand (SLIGKV). However, further deletion of the tethered ligand region (SLIGKV) at the N-terminus rescued the cell surface receptor expression and the response to the synthetic peptide ligand, suggesting that the signal peptide of PAR2 may be involved in preventing PAR2 from intracellular protease activation before reaching the cell surface. Supporting this hypothesis, an Arg36Ala mutation on PAR2ΔSP, which disabled the trypsin activation site, increased the receptor cell surface expression and the response to ligand stimulation. Similar effects were observed when PAR2ΔSP expressing cells were treated with protease inhibitors. Our findings indicated that these is a role of the PAR2 signal peptide in preventing the premature activation of PAR2 from intracellular protease cleavage before reaching the cells surface. The same mechanism may also apply to PAR1, PAR3, and PAR4.