Calcium-dependent Properties of Peptidylarginine Deiminase from Rabbit Skeletal Muscle

Abstract

Peptidylarginine deiminase, which catalyzes the deimination of arginyl residues in protein, required Ca2+ as an essential cofactor and the half-maximal activity was attained at 40~60 μM Ca2+. Other divalent cations were practically inactive except for Sr2+, which was about 50% as active as Ca2+ when tested at 10 Mm. However, Sr2+ at less than the concentration of 100 μM had little or no activity. The direct Ca2+-binding for the enzyme showed a sigmoidal curve with a transition midpoint of about 110 μM, indicating that the binding is cooperative. Analysis of Hill plots of the data revealed that the enzyme binds 3 mol of Ca2+ /mol of protein with an apparent dissociation constant of 110μM. A conformational change upon Ca2+-binding was also described for the enzyme using UV-difference spectra. The alteration could be attributed to an increased exposure of the aromatic residues to a more aqueous environment, as has been described for Ca2+-binding proteins such as calmodulin. Phosphatidylserine enhanced the reaction velocity and concomitantly reduced the Ca2+-requirement for the enzyme. These effects were stimulated by the addition of diacylglycerol. Diacylglycerol alone had little or no effect. On the other hand, calmodulin had no effect on the enzymatic activity over a wide range of Ca2+ concentrations. These suggest that the activity and Ca2+-sensitivity of peptidylarginine deiminase is increased at the cell membrane. © 1986, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.