Factor G utilizes a carbohydrate-binding cleft that is conserved between horseshoe crab and bacteria for the recognition of β-1,3-D-glucans

Yuki Ueda, Shuhei Ohwada, Yoshito Abe, Toshio Shibata, Manabu Iijima, Yukiko Yoshimitsu, Takumi Koshiba, Munehiro Nakata, Tadashi Ueda, Shun Ichiro Kawabata

研究成果: ジャーナルへの寄稿学術誌査読

13 被引用数 (Scopus)

抄録

In the horseshoe crab, the recognition of β-1,3-D-glucans by factor G triggers hemolymph coagulation. Factor G contains a domain of two tandem xylanase Z-like modules (Z1-Z2), each of which recognizes β-1,3-D-glucans. To gain an insight into the recognition of β-1,3-D-glucans from a structural view point, recombinants of Z1-Z2, the C-terminal module Z2, Z2 with a Cys to Ala substitution (Z2A), and its tandem repeat Z2A-Z2A were characterized. Z2 and Z1-Z2, but not Z2A and Z2A-Z2A, formed insoluble aggregates at higher concentrations more than ∼30 and 3 μM, respectively. Z1-Z2 and Z2A-Z2A bound more strongly to an insoluble β-1,3-D-glucan (curdlan) than Z2A. The affinity of Z2A for a soluble β-1,3-D-glucan (laminarin) was equivalent to those of Z1-Z2, Z2A-Z2A, and native factor G, suggesting that the binding of a single xylanase Z-like module prevents the subsequent binding of another module to laminarin. Interestingly, Z2A as well as intact factor G exhibited fungal agglutinating activity, and fungi were specifically detected with fluorescently tagged Z2A by microscopy. The chemical shift perturbation of Z2A induced by the interaction with laminaripentaose was analyzed by nuclear magnetic resonance spectroscopy. The ligand-binding site of Z2A was located in a cleft on a β-sheet in a predicted β-sandwich structure, which was superimposed onto cleft B in a cellulose-binding module of endoglucanase 5A from the soil bacterium Cellvibrio mixtus. We conclude that the pattern recognition for β-1,3-D-glucans by factor G is accomplished via a carbohydrate-binding cleft that is evolutionally conserved between horseshoe crab and bacteria.

本文言語英語
ページ(範囲)3810-3818
ページ数9
ジャーナルJournal of Immunology
183
6
DOI
出版ステータス出版済み - 9月 15 2009

!!!All Science Journal Classification (ASJC) codes

  • 免疫アレルギー学
  • 免疫学

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