Stratification of Chondroitin Sulfate Binding Sites in 3D-Model of Bovine Testicular Hyaluronidase and Effective Size of Glycosaminoglycan Coat of the Modified Protein

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A. V. Maksimenko^*, A. D. Turashev, and R. S. Beabealashvili

Institute of Experimental Cardiology, Russian Cardiology Research and Production Complex, 3-ya Cherepkovskaya ul. 15A, 121552 Moscow, Russia; fax: +7 (495) 414-6699; E-mail: alexmak@cardio.ru

^* To whom correspondence should be addressed.

Received February 21, 2014; Revision received September 29, 2014

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A 3D-model of bovine testicular hyaluronidase (BTH) was constructed based on established tertiary structure of human hyaluronidase Hyal1 using a molecular homological modeling method in silico. The analysis of the BTH 3D-model demonstrated lysine residue stratification during enzyme modification. The 3D-model of chondroitin sulfate (CHS)-modified hyaluronidase (BTH-CHS) was obtained by modeling covalent binding of lysine residues with benzoquinone-activated CHS. The degree of enzyme modification and the length of CHS chains were varied during 3D modeling. The importance of deep BTH modification degree for the formation of active and stable enzyme derivatives was shown, as determined earlier experimentally. The effective size of the CHS coat for productive BTH modification was confirmed. It is theoretically achieved at the increase in molecular mass of BTH-CHS derivative to approximately 140-180 kDa and can be practically obtained, according to experimental data, using CHS of different molecular mass (30-50 as well as 120-140 kDa).

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KEY WORDS: bovine testicular hyaluronidase, tertiary structure, chondroitin sulfate, lysine residue modification, hyaluronidase–chondroitin sulfate conjugate, modification degree, chondroitin sulfate coat

DOI: 10.1134/S0006297915030049

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