CC[O:3][C:4]([C@@H:6]([NH:14]C(C1C=CC=CC=1)=O)[CH2:7][CH2:8][CH2:9][N:10]=[C:11](N)[NH2:12])=[O:5].C1N(CC[OH:31])CCN(CCS(O)(=O)=O)C1.[Na+].[Cl-].C(C(O)=O)CP(CCC(O)=O)CCC(O)=O.[Cl-].[Cl-].[Ca+2]>>[NH2:14][C@H:6]([C:4]([OH:3])=[O:5])[CH2:7][CH2:8][CH2:9][NH:10][C:11]([NH2:12])=[O:31]Initial rates were determined in the absence and presence of various amounts of H3-amidine (0, 5, 10 mM), using BAEE as the substrate. BAEE and H3-amidine were pre-incubated in assay buffer containing 100 mM HEPES, pH 7.6, 50 mM NaCl, 0.5 mM TCEP, and 10 mM CaCl2 for 10 min at 37° C. Reactions were initiated by the addition of PAD4 to a final concentration of 0.2 μM. After incubation at 37° C. for 15 min, the reactions were quenched and the amount of Cit produced quantified. The initial rates derived from these experiments were fit by non-linear least fit squares to equations representing linear competitive inhibition (Equation 4) and linear noncompetitive inhibition (Equation 5), using the using the GraFit™ version 5.0.11 software package. v=Vm[S]/(Km(1+[I]/Kis)+[S]) (Eq. 4) v=Vm[S]I(Km(1+[I]/Kis)+[S](1+[I]/Kii)) (Eq. 5)
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