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Biochemistry (Moscow)

, Volume 75, Issue 9, pp 1173–1181 | Cite as

Destabilase-lysozyme of medicinal leech. Multifunctionality of recombinant protein

  • L. L. Zavalova
  • V. N. Lazarev
  • S. A. Levitsky
  • T. G. Yudina
  • I. P. BaskovaEmail author
Article

Abstract

Preparation and purification of a recombinant protein are described along with characteristics of its specific (for ɛ-(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-methylum-belliferyl-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-ɛ-(γ-Glu)-Lys- and exo-ɛ-(γ-Glu)-Lys-isopeptidolysis specific glycosidase activity 4-methylumbelliferyl-N-acetyl-β-chitotetraoxide muramidase activity antibacterial activity Methanosarcina barkeri 

Abbreviations

CM

cytoplasmic membrane

CW

cell wall

(GlcNAc)4-MeU

4-methylumbelliferryl-tetra-N-acetyl-β-chitotetraoxide

recDest-Lys

recombinant destabilase-lysozyme

SEM

scanning electron microscopy

TEM

transmission electron microscopy

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Copyright information

© Pleiades Publishing, Ltd. 2010

Authors and Affiliations

  • L. L. Zavalova
    • 1
  • V. N. Lazarev
    • 2
  • S. A. Levitsky
    • 2
  • T. G. Yudina
    • 3
  • I. P. Baskova
    • 3
    Email author
  1. 1.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Research Institute of Physicochemical MedicineMoscowRussia
  3. 3.Biological FacultyLomonosov Moscow State UniversityMoscowRussia

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