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Table 2.

Kinetic parameters determined for four arginine kinase (AK)constructs: Ensis full-length AK, D2—NusA—His-tag fusion protein, D2 alone and Limulus AKWTrev

ParameterEnsis full-lengthEnsis D2—NusA—His-tagEnsis D2Limulus WT
Arginine phosphate     
Kia (mmol l-10.333±0.046 0.618±0.083 0.592 (0.578, 0.606) 0.804±0.038 
Ka (mmol l-10.313±0.059 1.222±0.166 0.645 (0.625, 0.665) 0.226±0.004 
Kia/Ka 1.06 0.51 0.92 3.56 
ADP     
Kia (mmol l-10.050±0.008 0.087±0.012 0.076 (0.078, 0.074) 0.156±0.010 
Ka (mmol l-10.053±0.003 0.160±0.009 0.086 (0.084, 0.087) 0.043±0.002 
Kia/Ka 0.94 0.54 0.88 3.62 
Vmax (μmol min-1 mg-1196.65±17.41 (D2 only: 391.79) 175.57±7.45 (D2 only: 434.50) 446.07 (460.48, 431.65) 223.32±8.09 
Kcat (s-1266.31 295.33 303.21 156.32 
ParameterEnsis full-lengthEnsis D2—NusA—His-tagEnsis D2Limulus WT
Arginine phosphate     
Kia (mmol l-10.333±0.046 0.618±0.083 0.592 (0.578, 0.606) 0.804±0.038 
Ka (mmol l-10.313±0.059 1.222±0.166 0.645 (0.625, 0.665) 0.226±0.004 
Kia/Ka 1.06 0.51 0.92 3.56 
ADP     
Kia (mmol l-10.050±0.008 0.087±0.012 0.076 (0.078, 0.074) 0.156±0.010 
Ka (mmol l-10.053±0.003 0.160±0.009 0.086 (0.084, 0.087) 0.043±0.002 
Kia/Ka 0.94 0.54 0.88 3.62 
Vmax (μmol min-1 mg-1196.65±17.41 (D2 only: 391.79) 175.57±7.45 (D2 only: 434.50) 446.07 (460.48, 431.65) 223.32±8.09 
Kcat (s-1266.31 295.33 303.21 156.32 

With the exception of Ensis D2, all kinetic parameters are given as a mean value of at least three kinetic runs ± S.D. Ensis D2 values are given as a mean of two kinetic runs, with values from each run given in parentheses. Values shown in bold are Vmax values for Ensis full-length AK and D2—NusA—His-tag fusion protein if only the mass of D2 is taken into consideration.

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