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Interaction between human α2-macroglobulin and duodenase, a serine proteinase with dual specificity

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Abstract

Interaction between a serine proteinase from bovine duodenum and human serum α2-macroglobulin (α2-MG) was studied. α2-MG is established to be one of the most effective duodenase inhibitors. The enzyme is completely inhibited in less than 30 sec at equimolar ratio of the inhibitor and enzyme (concentration 2·10−8 M). Under identical conditions, the rate of duodenase association with α2-MG is at least 2.5-fold higher than the rate of chymotrypsin association with this inhibitor. The interaction with duodenase results in proteolysis of the inhibitor subunit in the “bait region”. Similarly to other proteases, duodenase in the complex with α2-MG retains the intact catalytic apparatus and ability to hydrolyze some small substrates. But the duodenase-inhibitor complex is fully inactive to proteins (bovine serum albumin). The stoichiometry of the enzyme interaction with the inhibitor is 2: 1 (mol/mol). Based on the association rate constant and the termination time of the duodenase and α2-MG in vivo association, α2-MG is suggested to be a physiological regulator of the enzyme.

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Abbreviations

α1-AT:

α1-inhibitor of proteinases (α1-antitrypsin)

α1-ACh:

α1-antichymotrypsin

α2-MG:

α2-macroglobulin

BAEE:

N-benzoyl-L-arginine ethyl ester

BBI:

soybean Baumann-Birk family proteinase inhibitor

DMSO:

dimethylsulfoxide

DTT:

dithiothreitol

PEG:

poly(ethylene glycol)

pNA:

p-nitroanilide

STI:

soybean Kunitz-type trypsin inhibitor

Suc:

N-succinyl

Tos:

tosyl

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Authors and Affiliations

  1. Department of Chemical Enzymology, Faculty of Chemistry, Lomonosov Moscow State University, 119992, Moscow, Russia

    O. O. Denisenko, N. A. Popykina & N. I. Larionova

  2. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    T. S. Zamolodchikova

Authors
  1. O. O. Denisenko
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  2. T. S. Zamolodchikova
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  3. N. A. Popykina
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  4. N. I. Larionova
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Corresponding author

Correspondence to N. I. Larionova.

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Original Russian Text © O. O. Denisenko, T. S. Zamolodchikova, N. A. Popykina, N. I. Larionova, 2006, published in Biokhimiya, 2006, Vol. 71, No. 6, pp. 815–824.

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Denisenko, O.O., Zamolodchikova, T.S., Popykina, N.A. et al. Interaction between human α2-macroglobulin and duodenase, a serine proteinase with dual specificity. Biochemistry (Moscow) 71, 658–666 (2006). https://doi.org/10.1134/S0006297906060101

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  • DOI: https://doi.org/10.1134/S0006297906060101

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