IBA Publikation
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DATUM, UNTERSCHRIFT / DATE, SIGNATURE
BANK AUSTRIA CREDITANSTALT, WIEN (IBAN AT04 1100 0006 2280 0100, BIC BKAUATWW), DEUTSCHE BANK MÜNCHEN (IBAN DE16 7007 0024 0238 8270 00, BIC DEUTDEDBMUC)
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IBA Publikation IBA Publikation
The Journal of Biological Chemistry 2009;284
Stephan Reitinger,
Johannes Müllegger,
Brigitte Greiderer,
Jens Erik Nielsen,
Günter Lepperdinger
S. 19173 - 19177 American Society for Biochemistry and Molecular Biology doi:10.1074/jbc.C109.004358
Abstract: Hyaluronidases from diverse species and sources have different pH optima. Distinct mechanisms with regard to dynamic structural changes, which control hyaluronidase activity at varying pH, are unknown. Human serum hyaluronidase 1 (HYAL1) is active solely below pH 5.1. Here we report the design of a HYAL1 variant that degrades hyaluronan up to pH 5.9. Besides highly conserved residues in close proximity of the active site of most hyaluronidases, we identified a bulky loop formation located at the end of the substrate binding crevice of HYAL1 to be crucial for substrate hydrolysis. The stretch between cysteine residues 207 and 221, which normally contains 13 amino acids, could be replaced by a tetrapeptide sequence of alternating glycine serine residues, thereby yielding an active enzyme with an extended binding cleft. This variant exhibited hyaluronan degradation at elevated pH. This is indicative for appropriate substrate binding and proper positioning being decisively affected by sites far off from the active center. Published Online: 2009/11/05 14:06:16 Object Identifier: 0xc1aa5576 0x0022a4ea Rights: .
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epub.oeaw – Institutionelles Repositorium der Österreichischen Akademie der Wissenschaften epub.oeaw – Institutional Repository of the Austrian Academy of Sciences
A-1011 Wien, Dr. Ignaz Seipel-Platz 2
Tel. +43-1-515 81/DW 3420, Fax +43-1-515 81/DW 3400 http://epub.oeaw.ac.at, e-mail: epub@oeaw.ac.at |