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J. Biol. Chem., Vol. 279, Issue 24, 25307-25312, June 11, 2004

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Human Mannose-binding Lectin and L-Ficolin Function as Specific Pattern Recognition Proteins in the Lectin Activation Pathway of Complement^*

Young Gerl Ma, Mi Yhang Cho, Mingyi Zhao, Ji Won Park, Misao Matsushita, Teizo Fujita¶, and Bok Luel Lee||

From the College of Pharmacy, Pusan National University, Jangjeon Dong, Kumjeong Ku, Busan 609-735, Korea, Institute of Glycotechnology and Department of Applied Biochemistry, Tokai University, 1117-Kitakaname, Hiratsuka 259-1292, Japan, ^¶Department of Biochemistry, Fukushima Medical University School of Medicine, 1-Hikarigao-ka, and CREST, Japan Science and Technology Agency, Fukushima 960-1295, Japan

The innate immune response in vertebrates and invertebrates requires the presence of pattern recognition receptors or proteins that recognize microbial cell components including lipopolysaccharide, bacterial peptidoglycan (PGN), and fungal 1,3--D-glucan. We reported previously that PGN and 1,3--D-glucan recognition proteins from insect hemolymph were able to induce the activation of the prophenoloxidase-activating system, one of the major invertebrate innate immune reactions. The goal of this study was to characterize the biochemical properties and effects of the human counterparts of these molecules. Soluble pattern recognition proteins were purified from human serum and identified as human mannose-binding lectin (MBL) and L-ficolin. The use of specific microbial cell component-coupled columns demonstrated that MBL and L-ficolin bind to PGN and 1,3--D-glucan, respectively. Purified MBL and L-ficolin were associated with MBL-associated serine proteases-1 and -2 (MASPs) and small MBL-associated protein as determined by Western blot analysis. Finally, the binding of purified MBL/MASP and L-ficolin/MASP complexes to PGN and 1,3--D-glucan, respectively, resulted in the activation of the lectin-complement pathway. These results indicate that human PGN and 1,3--D-glucan recognition proteins function as complement-activating lectins.

Received for publication, January 21, 2004 , and in revised form, March 27, 2004.

^* This work was supported in part by a research grant from the Korean Research Foundation (KRF-2003-041-C20203), a Korea Food and Drug Administration grant (KFDA-03142-BIO-022) (to B. L. L.), and a Pusan National University Research grant (to B. L. L.). 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.

^|| To whom correspondence should be addressed. Tel.: 82-51-510-2809; Fax: 82-51-513-6754; E-mail: brlee{at}pusan.ac.kr.

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