Activated protein C (APC) reduces mortality in severe sepsis patients and exhibits beneficial effects in multiple animal injury models. APC anticoagulant activity involves inactivation of factors Va and VIIIa whereas APC cytoprotective activities involve the endothelial protein C receptor (EPCR) and protease-activated receptor-1 (PAR-1). The relative importance of APC’s anticoagulant activity vs. APC’s direct cytoprotective effects on cells for the in vivo benefits is unclear. To distinguish cytoprotective from anticoagulant activities of APC, a protease domain mutant, 5A-APC (RR229/230AA and KKK191-193AAA), was made and compared to recombinant wild type (rwt)-APC. This mutant had minimal anticoagulant activity but normal cytoprotective activities that were dependent on EPCR and PAR-1. Whereas anticoagulantly active rwt-APC inhibited secondary-extended thrombin generation and concomitant thrombin-dependent activation of thrombin activatable fibrinolysis inhibitor (TAFI) in plasma, secondary-extended thrombin generation and the activation of TAFI was essentially unopposed by 5A-APC due to its low anticoagulant activity. Compared to rwt-APC, 5A-APC had minimal profibrinolytic activity and preserved TAFI-mediated anti-inflammatory carboxypeptidase activities towards bradykinin and presumably towards the anaphlatoxins, C3a and C5a, which are well known pathological mediators in sepsis. Thus, genetic engineering can selectively alter APC's multiple activities and provide APC mutants that retain APC’s beneficial cytoprotective effects while diminishing bleeding risk due to reduction in APC's anticoagulant and APC-dependent profibrinolytic activities.