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J. Biol. Chem., Vol. 283, Issue 27, 18530-18537, July 4, 2008

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The Transfer of Heavy Chains from Bikunin Proteins to Hyaluronan Requires Both TSG-6 and HC2^*

Kristian W. Sanggaard, Carsten S. Sonne-Schmidt, Toke P. Krogager, Karen A. Lorentzen, Hans-Georg Wisniewski, Ida B. Thøgersen, and Jan J. Enghild¹

From the Center for Insoluble Protein Structures (inSPIN) and Interdisciplinary Nanoscience Center (iNANO) at the Department of Molecular Biology, Science Park, University of Aarhus, Gustav Wiedsvej 10C, 8000 Aarhus C, Denmark and Department of Microbiology, New York University School of Medicine, New York, New York 10016

Tumor necrosis factor-stimulated gene-6 protein (TSG-6) is involved in the transfer of heavy chains (HCs) from inter--inhibitor (II), pre--inhibitor, and as shown here HC2·bikunin to hyaluronan through the formation of covalent HC·TSG-6 intermediates. In contrast to II and HC2·bikunin, pre--inhibitor does not form a covalent complex in vitro using purified proteins but needs the presence of another factor (Rugg, M. S., Willis, A. C., Mukhopadhyay, D., Hascall, V. C., Fries, E., Fülöp, C., Milner, C. M., and Day, A. J. (2005) J. Biol. Chem. 280, 25674–25686). In the present study we purified the required component from human plasma and identified it as HC2. Proteins containing HC2 including II, HC2·bikunin, and free HC2 promoted the formation of HC3·TSG-6 and subsequently HC3·hyaluronan complexes. HC1 or HC3 did not possess this activity. The presented data reveal that both HC2 and TSG-6 are required for the transesterification reactions to occur.

Received for publication, February 1, 2008 , and in revised form, April 10, 2008.

^* This work was supported by the Danish National Research Foundation (to J. J. E.), the Danish Natural Science Research Council (to J. J. E.), and the Carlsberg Foundation (to K. W. S.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Tables S1–S6 and Figs. S1 and S2.

¹ To whom correspondence should be addressed. Tel.: 45-8942-5062; Fax: 45-8942-5063; E-mail: jje{at}mb.au.dk.

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