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Isolation and Characterization of the ALP1 Protease from Aspergillus fumigatus and Its Protein Inhibitor from Physarium polycephalum

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Abstract

It is known that Aspergillus fumigatus secretes a serine protease ALP1 of the subtilisin family in the presence of extracellular protein substrates. We found conditions of A. fumigatus culturing that provide a high ALP1 activity inside cells without induction by extracellular proteins. The identity of the properties of the secreted and intracellular enzymes was shown. A thermostable protein inhibitor of the ALP1 protease was isolated from the plasmodium of myxomycete Physarum polycephalum. Its molecular mass is 32–33 kDa. It inhibits the ALP1 protease activity with IC50 of 0.14 μM and was also shown to be a less efficient inhibitor of the activity of HIV-1 protease (IC50 2.5 μM).

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Abbreviations

Azocoll:

azocollagen from Sigma, United States

EGTA:

ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

HIV:

human immunodeficiency virus

MES:

2-(N-morpholino) ethanesulfonic acid

PMSF:

phenylmethylsulfonyl fluoride

TLCK:

Nα-tosyl-L-lysine chloromethyl ketone hydrochloride

TPCK:

L-1-chloro-3-(4-tosylamido)-4-phenyl-2-butanone

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Authors and Affiliations

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

    N. A. Kalinina, L. V. Samokhvalova, G. V. Malakhova, A. M. Volynskaya & V. A. Nesmeyanov

  2. London Hospitals, Department of Clinical Microbiology, University College, Grafton Way, London, United Kingdom

    D. A. Davies, G. Scott & G. Venning

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  1. D. A. Davies
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  2. N. A. Kalinina
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  3. L. V. Samokhvalova
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  4. G. V. Malakhova
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  5. G. Scott
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  7. A. M. Volynskaya
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  8. V. A. Nesmeyanov
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__________

Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 3, 2005, pp. 259–268.

Original Russian Text Copyright © 2005 by Davies, Kalinina, Samokhvalova, Malakhova, Scott, Venning, Volynskaya, Nesmeyanov.

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Davies, D.A., Kalinina, N.A., Samokhvalova, L.V. et al. Isolation and Characterization of the ALP1 Protease from Aspergillus fumigatus and Its Protein Inhibitor from Physarium polycephalum. Russ J Bioorg Chem 31, 229–237 (2005). https://doi.org/10.1007/s11171-005-0032-6

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  • DOI: https://doi.org/10.1007/s11171-005-0032-6

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