Induction of Catalytic Activity of Plasminogen by Monoclonal Antibody IV-Ic in the Presence of Divalent Metal Cations and alpha₂-Antiplasmin

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L. I. Sokolovskaya^*, A. Yu. Slominskii, and G. L. Volkov

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, ul. Leontovicha 9, Kiev 01030, Ukraine; fax: (38-044) 235-5172; E-mail: sokludmila@yandex.ru

^* To whom correspondence should be addressed.

Received July 21, 2005; Revision received October 21, 2005

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Investigation of the influence of divalent metal cations on the induction of plasminogen catalytic activity by monoclonal antibody IV-Ic showed that the presence of metal cations in the reaction medium changes the induction by slowing down or accelerating the process. Ions of Zn²⁺, Mn²⁺, and Cu²⁺ completely inhibit activation. Ions of Co²⁺ and Ni²⁺ decrease the rate of the first and second phases of the reaction more than 2 times. Ca²⁺ ions do not have any effect on the activation rate. Ions of Mg²⁺, Ba²⁺, and Sr²⁺ increase the rate of the first phase of the reaction by 1.5, 2.0, and 2.0 times and the rate of the second phase by 2.0, 3.8, and 4.7 times, correspondingly. Sr²⁺ ions have the strongest stimulating effect on plasminogen activation by monoclonal antibody IV-Ic. Investigation of the dose dependent effect of Sr²⁺ on the rate of plasminogen activation by monoclonal antibody IV-Ic showed stimulating effect of Sr²⁺ at concentrations from 0.1 to 1.0 mM with half maximum at 0.6 mM. However, Sr²⁺ ions do not affect amidolytic activity of plasmin and activation of plasminogen by streptokinase. Sr²⁺ ions also do not affect monoclonal antibody IV-Ic binding to plasminogen. The effect of Sr²⁺ is specific and mediated by the IV-Ic component. The presence of metal cations affects conformational changes in the process of active site formation. Metal cations also affect structure of the plasminogen molecule active site in the complex with monoclonal antibody IV-Ic and enzyme-substrate interaction. The effect of alpha₂-antiplasmin on the induction of plasminogen catalytic activity by monoclonal antibody IV-Ic in range of concentrations from 5 to 30 nM has been studied. alpha₂-Antiplasmin at concentration 30 nM almost completely inhibits induction of plasminogen catalytic activity by monoclonal antibody IV-Ic at the ratio plasminogen/alpha₂-antiplasmin of 3 : 1. This can be explained by competition of alpha₂-antiplasmin and monoclonal antibody IV-Ic for the lysine-binding sites of plasminogen and inhibition of the active center in activated complex plasminogen*-mAB IV-Ic. Divalent metal cations and alpha₂-antiplasmin are important factors in induction of plasminogen catalytic activity by monoclonal antibody IV-Ic.

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KEY WORDS: plasminogen, monoclonal antibody, divalent metal cations, alpha₂-antiplasmin

DOI: 10.1134/S000629790606006X

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