Crystallization and preliminary X-ray diffraction study of the protealysin precursor belonging to the peptidase family M
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CTURE OF MACROMOLECULAR COMPOUNDS
Crystallization and Preliminary X-Ray Diffraction Study of the Protealysin Precursor Belonging to the Peptidase Family M4 T. Yu. Gromovaa, b, I. V. Demidyuka, S. V. Kostrova, N. I. Sosfenovc, V. R. Melik-Adamyanc, and I. P. Kuranovac a
Institute of Molecular Genetics, Russian Academy of Sciences, pl. Kurchatova 2, Moscow, 123182 Russia e-mail: [email protected] b Moscow State Academy of Fine Chemical Technology, pr. Vernadskogo 86, Moscow, 119571 Russia e-mail: [email protected] c Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninskiœ pr. 59, Moscow, 119333 Russia e-mail: [email protected] Received December 26, 2007
Abstract—A protealysin precursor (the enzyme of the peptidase family M4) was crystallized for the first time. The crystal-growth conditions were found, and single crystals of the protein with dimensions of 0.3–0.5 mm were grown. The preliminary X-ray diffraction study of the enzyme was performed. The protealysin precursor was shown to crystallize in two crystal modifications suitable for the X-ray diffraction study of the three-dimensional structure of the protein molecule at atomic resolution. PACS numbers: 87.15.nt DOI: 10.1134/S1063774508050118
INTRODUCTION Thermolysin-like proteinases (TLPs) or peptidases of the family M4 are a well-studied group of enzymes. Proteins of this family were found in numerous grampositive and gram-negative bacteria. In addition, TLPs were found in fungi and the Methanosarcina acetivorans archaebacterium [1]. Peptidases of the family M4 contain a zinc ion in the catalytic site [2] and are prone to hydrolysis of peptide bonds formed by amino groups of bulky hydrophobic amino-acid residue [3]. Three-dimensional structures of several representatives of this family were established: thermolysin [4–7] and proteinases from Bacillus cereus [8, 9], Pseudomonas aeruginosa [10], and Staphylococcus aureus [11]. Thermolysin-like proteinases, like many other proteolytic enzymes [12], are synthesized as precursors whose polypeptide chains involve additional structural elements which are absent in the mature molecules. It has been demonstrated [13] that peptidases of the family M4 can be classified into two groups according to the structures of the amino-terminal domains (ATDs) of precursors that are removed during maturation. Wellknown TLPs, including the above-mentioned thermolysin and proteinases from B. cereus, P. aeruginosa, and S. aureus, belong to one group. The precursors of these proteins include ATDs with a length of up to 200 amino-acid residues (long ATDs). Long ATDs consist of a signal peptide that is responsible for the secre-
tion and a following propeptide. Propeptides play a very important role in the action of TLPs containing long ATDs. These structural elements serve as intramolecular chaperones, which are generally necessary for the formation of mature active proteinases [14–19]. In addition, propeptides act as inhibitors of mature enzymes [17–21] and are apparently involved in the secretion of the correspo
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