Arginase

I.U.B.: 3.5.3.1 L-Arginine amidinohydrolase Enzymatic Reaction (image will open in a new window) Arginase catalyzes the following reaction:

Molecular weight: 115,000-120,000 (Harell and Sokolovsky 1972). Composition: The amino acid content has been reported by Greenberg (1960). According to Harell and Sokolovsky (1972) the enzyme does not split into subunits; it binds 4 atoms of Mn2+which are essential to its activity as well as stability and has a hexose content of 3-5%. Optimum pH: 10 (Mn2+ activated, Greenberg 1960). Extinction coefficient: = 9.6 (Harell and Sokolovsky 1972).

Activators: Mn2+, Ni2+, or Co2+. Specificity: Arginase activity requires the free guanidino group and the free carboxyl group of arginine (Greenberg 1960). Inhibitors: Hg2+, Ag2+, Zn2+. Stability: Bond (1973) reports that susceptibility of beef liver arginase to proteolytic inactivation is enhanced by increasing activating concentration of Mn2+. Cysteine, isoleucine, leucine, valine and alanine protect the enzyme against trypsin inactivation. The enzyme is highly stable when stored refrigerated as a solution at neutral pH or as a lyophilized powder.

Calculation Determine the micromoles of urea released from a standard urea curve in the range of 0.1-1.0 micromoles urea.

1 unit=1 u mol of ornithine formed per mg arginase per min -1.

Molar absorptivities (= molar extinction coefficients) for many proteins are provided in the Practical Handbook ofBiochemistry and Molecular Biology.3 Expressed in this form, the extinction coefficient allows for estimation of the molarconcentration of a solution from its measured absorbance. A / = molar concentration Extinction coefficients for proteins are generally reported with respect to an absorbance measured at or near a wavelength of 280 nm. Although the absorption maxima for certain proteins may be at other wavelengths, 280 nm is favored because proteins absorb strongly there while other substances commonly in protein solutions do not.

Proteins and Protein Mixtures with Unknown Extinction Coefficients If no extinction coefficient information exists for a protein or protein mixture of interest, and a rough estimate of protein concentration is required for a solution that has no other interfering substances, assume percent = 10. Most protein extinction coefficients ( percent) fall in the range 4.0-24.0.3 Therefore, although any given protein can vary significantly from percent = 10, the average for a mixture of many different proteins will likely be close to 10.

Extinction Coefficient at 515nm,25mM concentration : 20600.0