HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 282, Issue 8, 5597-5607, February 23, 2007

This Article Full Text Full Text (PDF) All Versions of this Article: 282/8/5597    most recent M608358200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Harada, H. Articles by Takahashi, M. Search for Related Content PubMed PubMed Citation Articles by Harada, H. Articles by Takahashi, M. Social Bookmarking                      What's this?

CD44-dependent Intracellular and Extracellular Catabolism of Hyaluronic Acid by Hyaluronidase-1 and -2^*

From the Lead Discovery Research Laboratories, Sankyo Co., Ltd., 1-2-58 Hiromachi, Shinagawa-ku, Tokyo 140-8710, Japan

Hyaluronic acid (HA) is a high molecular weight glycosaminoglycan involved in a wide variety of cellular functions. However, its turnover in living cells remains largely unknown. In this study, CD44, a receptor for HA, and hyaluronidase-1, -2, and -3 (Hyal-1, -2 and -3) were stably expressed in HEK 293 cells and the mechanism of HA catabolism was systematically investigated using fluorescein-labeled HA. CD44 was essential for HA degradation by both endogenous and exogenously expressed hyaluronidases. Hyal-1 was not able to cleave HA in living cells in the absence of CD44. Intracellular HA degradation was predominantly mediated by Hyal-1 after incorporation of HA by CD44. Although Hyal-1 was active only in intracellular space in vivo, a certain amount of the enzyme was secreted to extracellular space. This extracellular Hyal-1 was found to be incorporated by cells and such uptake of Hyal-1 was, in part, involved in the intracellular degradation of HA. Hyal-2 was involved in the extracellular degradation of HA. Hyal-2 activity was also dependent on the expression of CD44 in both living cells and enzyme assays. Immunofluorescent microscopy demonstrated that both Hyal-2 and CD44 are present on the cell surface. Without CD44 expression, Hyal-2 existed in a granular pattern, and did not show hyaluronidase activity, suggesting that localization change could contribute to Hyal-2 function. A convenient and quantitative enzyme assay was established for the measurement of Hyal-2 activity. Hyal-2 activity was detected in the membrane fraction of cells co-expressing Hyal-2 and CD44. The pH optimum for Hyal-2 was 6.0–7.0. The membrane fraction of cells expressing Hyal-2 alone did not show hyaluronidase activity. Hyal-3 did not show any hyaluronidase activity in our experimental conditions. Based on these findings, Hyal-1 and -2 contribute to intracellular and extracellular catabolism of HA, respectively, in a CD44-dependent manner, and their HA degradation occurs independently from one another.

Received for publication, August 31, 2006 , and in revised form, November 30, 2006.

^* 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.

¹ Present address: Tsukuba Research Institute, Banyu Pharmaceutical Co., Ltd., 3 Okubo, Tsukuba, Ibaraki 300-2611, Japan. E-mail: hosami_harada{at}merck.com.

² To whom correspondence should be addressed. Tel.: 81-3-3492-3131; Fax: 81-3-5436-8578; E-mail: mtakah{at}sankyo.co.jp.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				D. Kothapalli, J. Flowers, T. Xu, E. Pure, and R. K. Assoian Differential Activation of ERK and Rac Mediates the Proliferative and Anti-proliferative Effects of Hyaluronan and CD44 J. Biol. Chem., November 14, 2008; 283(46): 31823 - 31829. [Abstract] [Full Text] [PDF]

				A. M. Orimoto, K. Dumaresq-Doiron, J.-Y. Jiang, N. Tanphaichitr, B. K. Tsang, and E. Carmona Mammalian Hyaluronidase Induces Ovarian Granulosa Cell Apoptosis and Is Involved in Follicular Atresia Endocrinology, November 1, 2008; 149(11): 5835 - 5847. [Abstract] [Full Text] [PDF]

				M. E. Monzon, D. Manzanares, N. Schmid, S. M. Casalino-Matsuda, and R. M. Forteza Hyaluronidase Expression and Activity Is Regulated by Pro-Inflammatory Cytokines in Human Airway Epithelial Cells Am. J. Respir. Cell Mol. Biol., September 1, 2008; 39(3): 289 - 295. [Abstract] [Full Text] [PDF]

				N. Itano Simple Primary Structure, Complex Turnover Regulation and Multiple Roles of Hyaluronan J. Biochem., August 1, 2008; 144(2): 131 - 137. [Abstract] [Full Text] [PDF]

				D. C. Martin, V. Atmuri, R. J. Hemming, J. Farley, J. S. Mort, S. Byers, S. Hombach-Klonisch, R. Stern, and B. L. Triggs-Raine A mouse model of human mucopolysaccharidosis IX exhibits osteoarthritis Hum. Mol. Genet., July 1, 2008; 17(13): 1904 - 1915. [Abstract] [Full Text] [PDF]

				R. Hemming, D. C. Martin, E. Slominski, J. I. Nagy, A. J. Halayko, S. Pind, and B. Triggs-Raine Mouse Hyal3 encodes a 45- to 56-kDa glycoprotein whose overexpression increases hyaluronidase 1 activity in cultured cells Glycobiology, April 1, 2008; 18(4): 280 - 289. [Abstract] [Full Text] [PDF]

				D. Vigetti, M. Viola, E. Karousou, M. Rizzi, P. Moretto, A. Genasetti, M. Clerici, V. C. Hascall, G. De Luca, and A. Passi Hyaluronan-CD44-ERK1/2 Regulate Human Aortic Smooth Muscle Cell Motility during Aging J. Biol. Chem., February 15, 2008; 283(7): 4448 - 4458. [Abstract] [Full Text] [PDF]

				A. G. Bharadwaj, K. Rector, and M. A. Simpson Inducible Hyaluronan Production Reveals Differential Effects on Prostate Tumor Cell Growth and Tumor Angiogenesis J. Biol. Chem., July 13, 2007; 282(28): 20561 - 20572. [Abstract] [Full Text] [PDF]