Thrombin is known to signal to cells by cleaving/activating a G-protein-coupled family of proteinase-activated receptors (PARs). The signaling mechanism involves the proteolytic unmasking of an N-terminal receptor sequence that acts as a 'tethered' receptor-activating ligand (TL). To date, the recognized targets of thrombin cleavage-activation for signaling are PARs 1 and 4, in which thrombin cleaves at a conserved target arginine to reveal a tethered ligand. PAR2, which like PAR1 is also cleaved at an N-terminal arginine to unmask its tethered-ligand, is generally regarded as a target for trypsin but not for thrombin signaling. We now show that thrombin, at concentrations that can be achieved at sites of acute injury or in a tumor microenvironment, can directly activate PAR2 vasorelaxation and signaling, stimulating calcium and MAPKinase responses along with triggering beta-arrestin recruitment. Thus, PAR2 can be added alongside PARs 1 and 4 to the targets whereby thrombin can affect tissue function.