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Kinetics of chaperone activity of proteins Hsp70 and Hdj1 in human leukemia U-937 cells after preconditioning with thermal shock or compound U-133

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Abstract

Kinetics of the chaperone activity of proteins Hsp70 and Hdj1 were analyzed in human U-937 promonocytes during their response to heat shock or to treatment with the echinochrome triacetyl glucoside derivative U-133. To measure the chaperone activity of both proteins, a special test was developed for their recognition and binding of a denatured protein. Using this test, the chaperone activity could be concurrently estimated in large numbers of cellular or tissue extracts. We also estimated the contents of both chaperones in cells by immunoblotting. The values for contents of Hsp70 and Hdj1 obtained by two independent test systems coincided, and this suggested that the substrate-binding activity could change proportionally to the chaperone content in the protein mixture. Therefore, the test developed by us can be employed for high throughput screening of drugs activating cellular chaperones. The analysis of quantity and activity of two cellular chaperones during the cell response to heat stress or to the drug-like substance U-133 showed that both factors caused the accumulation of chaperones with similar kinetics. We conclude that the efficiency of drug preconditioning could be close to the efficiency of hyperthermia and that the high activity of chaperones could be retained in human cells for no less than 1.5 days.

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Abbreviations

CMLA:

carboxymethylated lactalbumin

DTT:

dithiothreitol

EIA:

enzyme immunoassay

PBS:

phosphatebuffered saline

U-103:

tetra-O-acetyl-β-D-glucopyranoside

U-133:

echinochrome triacetyl glucoside

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Authors and Affiliations

  1. Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, 194064, St. Petersburg, Russia

    V. F. Lazarev, K. V. Onokhin, O. I. Antimonova, I. V. Guzhova & B. A. Margulis

  2. Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian Academy of Sciences, pr. 100 let Vladivostoku 159, 690022, Vladivostok, Russia

    S. G. Polonik

Authors
  1. V. F. Lazarev
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  2. K. V. Onokhin
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  3. O. I. Antimonova
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  4. S. G. Polonik
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  5. I. V. Guzhova
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  6. B. A. Margulis
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Corresponding author

Correspondence to B. A. Margulis.

Additional information

Original Russian Text © V. F. Lazarev, K. V. Onokhin, O. I. Antimonova, S. G. Polonik, I. V. Guzhova, B. A. Margulis, 2011, published in Biokhimiya, 2011, Vol. 76, No. 5, pp. 724–730.

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Lazarev, V.F., Onokhin, K.V., Antimonova, O.I. et al. Kinetics of chaperone activity of proteins Hsp70 and Hdj1 in human leukemia U-937 cells after preconditioning with thermal shock or compound U-133. Biochemistry Moscow 76, 590–595 (2011). https://doi.org/10.1134/S0006297911050099

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  • DOI: https://doi.org/10.1134/S0006297911050099

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