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Destabilase-Lysozyme of Medicinal Leech. Multifunctionality of Recombinant Protein

L. L. Zavalova1, V. N. Lazarev2, S. A. Levitsky2, T. G. Yudina3, and I. P. Baskova3*

1Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 119997 Moscow, Russia; fax: (495) 330-6538; E-mail: lz@ibch.ru

2Research Institute of Physicochemical Medicine, ul. Malaya Pirogovskaya 1a, 119992 Moscow, Russia; fax: (499) 255-2846; E-mail: lazar0@newmail.ru

3Biological Faculty, Lomonosov Moscow State University, 119991 Moscow, Russia; fax: (495) 939-4309; E-mail: Saliva1@yandex.ru

* To whom correspondence should be addressed.

Received February 27, 2010; Revision received March 27, 2010
Preparation and purification of a recombinant protein are described along with characteristics of its specific (for ε-(γ-Glu)-Lys and D-dimer substrates) and nonspecific (for L-γ-Glu-pNA) isopeptidase activities; the absence of peptidase function for α-(α-Glu)-Lys substrate is noted. It is shown that the protein exhibits muramidase (cell walls of Micrococcus lysodeikticus) and specific glycosidase activities. The latter was determined towards the fluorogenic substrate 4-methylumbelliferyl-tetra-N-acetyl-β-chitotetraoxide. Antimicrobial activity of recombinant destabilase-lysozyme protein (recDest-Lys) and its 11-membered amphipathic peptide was revealed towards cells of the strict anaerobic Archaean Methanosarcina barkeri, whose cell walls contain no murein. Possible mechanisms of the effect of recDest-Lys on these cells are discussed.
KEY WORDS: endo-ε-(γ-Glu)-Lys- and exo-ε-(γ-Glu)-Lys-isopeptidolysis, specific glycosidase activity, 4-methylumbelliferyl-N-acetyl-β-chitotetraoxide, muramidase activity, antibacterial activity, Methanosarcina barkeri

DOI: 10.1134/S0006297910090129