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Microbe-specific C3b deposition in the horseshoe crab complement system in a C2/factor B-dependent or -independent manner.

Tagawa K, Yoshihara T, Shibata T, Kitazaki K, Endo Y, Fujita T, Koshiba T, Kawabata S - PLoS ONE (2012)

Bottom Line: Complement C3 plays an essential role in the opsonization of pathogens in the mammalian complement system, whereas the molecular mechanism underlying C3 activation in invertebrates remains unknown.TtC2/Bf-1 and TtC2/Bf-2 were synthesized and glycosylated in hemocytes and secreted to hemolymph plasma, which existed in a complex with C3 (TtC3), and their activation by microbes was absolutely Mg(2+)-dependent.We conclude that plasma lectins and factor C play key roles in microbe-specific TtC3b deposition in a C2/factor B-dependent or -independent manner.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Systems Life Sciences, Kyushu University, Fukuoka, Japan.

ABSTRACT
Complement C3 plays an essential role in the opsonization of pathogens in the mammalian complement system, whereas the molecular mechanism underlying C3 activation in invertebrates remains unknown. To understand the molecular mechanism of C3b deposition on microbes, we characterized two types of C2/factor B homologs (designated TtC2/Bf-1 and TtC2/Bf-2) identified from the horseshoe crab Tachypleus tridentatus. Although the domain architectures of TtC2/Bf-1 and TtC2/Bf-2 were identical to those of mammalian homologs, they contained five-repeated and seven-repeated complement control protein domains at their N-terminal regions, respectively. TtC2/Bf-1 and TtC2/Bf-2 were synthesized and glycosylated in hemocytes and secreted to hemolymph plasma, which existed in a complex with C3 (TtC3), and their activation by microbes was absolutely Mg(2+)-dependent. Flow cytometric analysis revealed that TtC3b deposition was Mg(2+)-dependent on Gram-positive bacteria or fungi, but not on Gram-negative bacteria. Moreover, this analysis demonstrated that Ca(2+)-dependent lectins (C-reactive protein-1 and tachylectin-5A) were required for TtC3b deposition on Gram-positive bacteria, and that a Ca(2+)-independent lectin (Tachypleus plasma lectin-1) was definitely indispensable for TtC3b deposition on fungi. In contrast, a horseshoe crab lipopolysaccharide-sensitive protease factor C was necessary and sufficient to deposit TtC3b on Gram-negative bacteria. We conclude that plasma lectins and factor C play key roles in microbe-specific TtC3b deposition in a C2/factor B-dependent or -independent manner.

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Proposed mechanism for deposition of TtC3b on the surfaces of microbes.On the surface of Gram-negative bacteria, the LPS-sensitive protease factor C is activated through the interaction with LPS. Activated factor C then acts as a C3 convertase, and the resulting TtC3b is deposited. On the Gram-positive bacteria, Ca2+-dependent lectins such as TtCRP-1 (Tachypleus tridentatus C-reactive protein-1) and TL-5A (Tachylectin-5A) recruit the complex (C3-B), TtC3-TtC2/Bf-1, or TtC3-TtC2/Bf-2. The second C3 convertase (C3b-Bb), TtC3b-TtC2/Bf-1b or TtC3b-TtC2/Bf-2b complex, may be formed by an unidentified protease on the surfaces of Gram-positive bacteria and fungi. TL-1 (Tachylectin-1) and possibly TPL-1 (Tachypleus plasma lectin-1) also promote TtC3b deposition on the three types of microbes, especially on fungi.
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pone-0036783-g009: Proposed mechanism for deposition of TtC3b on the surfaces of microbes.On the surface of Gram-negative bacteria, the LPS-sensitive protease factor C is activated through the interaction with LPS. Activated factor C then acts as a C3 convertase, and the resulting TtC3b is deposited. On the Gram-positive bacteria, Ca2+-dependent lectins such as TtCRP-1 (Tachypleus tridentatus C-reactive protein-1) and TL-5A (Tachylectin-5A) recruit the complex (C3-B), TtC3-TtC2/Bf-1, or TtC3-TtC2/Bf-2. The second C3 convertase (C3b-Bb), TtC3b-TtC2/Bf-1b or TtC3b-TtC2/Bf-2b complex, may be formed by an unidentified protease on the surfaces of Gram-positive bacteria and fungi. TL-1 (Tachylectin-1) and possibly TPL-1 (Tachypleus plasma lectin-1) also promote TtC3b deposition on the three types of microbes, especially on fungi.

Mentions: Figure 9 shows a schematic model of TtC3b deposition on the surfaces of three types of microbes. The Ca2+-dependent lectins (TtCRP-1 and TL-5A) and the Ca2+-independent lectin (TLP-1/TL-1) seem to recruit the complex between TtC3 and TtC2/Bf-1 or TtC2/Bf-2 (C3(H2O)B) on Gram-positive bacteria and fungi, respectively, to convert the complex to a TtC3 convertase (C3bBb). These lectins seem to work as carries for the C3bBb-like convertase to the surface of microbes with quite specific binding activities. However, these lectins have broad binding specificities against microbial cell wall substances and bind not only to Gram-positive bacteria but also to Gram-negative bacteria [7], [9], [10], [13], [14]. In addition, TPL-1 and TL-1 bind to saccharides, such as N-acetyl-glucosamine, galactose, and mannan [13], [14], and possibly bind to β-1,3-D-glucan, a major cell wall component of fungi. Therefore, the binding specificities of these lectins are not sufficient to explain the microbe specific TtC3b deposition. Unknown proteins may be additionally required to determine the specific target of these lectins.


Microbe-specific C3b deposition in the horseshoe crab complement system in a C2/factor B-dependent or -independent manner.

Tagawa K, Yoshihara T, Shibata T, Kitazaki K, Endo Y, Fujita T, Koshiba T, Kawabata S - PLoS ONE (2012)

Proposed mechanism for deposition of TtC3b on the surfaces of microbes.On the surface of Gram-negative bacteria, the LPS-sensitive protease factor C is activated through the interaction with LPS. Activated factor C then acts as a C3 convertase, and the resulting TtC3b is deposited. On the Gram-positive bacteria, Ca2+-dependent lectins such as TtCRP-1 (Tachypleus tridentatus C-reactive protein-1) and TL-5A (Tachylectin-5A) recruit the complex (C3-B), TtC3-TtC2/Bf-1, or TtC3-TtC2/Bf-2. The second C3 convertase (C3b-Bb), TtC3b-TtC2/Bf-1b or TtC3b-TtC2/Bf-2b complex, may be formed by an unidentified protease on the surfaces of Gram-positive bacteria and fungi. TL-1 (Tachylectin-1) and possibly TPL-1 (Tachypleus plasma lectin-1) also promote TtC3b deposition on the three types of microbes, especially on fungi.
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Related In: Results  -  Collection

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pone-0036783-g009: Proposed mechanism for deposition of TtC3b on the surfaces of microbes.On the surface of Gram-negative bacteria, the LPS-sensitive protease factor C is activated through the interaction with LPS. Activated factor C then acts as a C3 convertase, and the resulting TtC3b is deposited. On the Gram-positive bacteria, Ca2+-dependent lectins such as TtCRP-1 (Tachypleus tridentatus C-reactive protein-1) and TL-5A (Tachylectin-5A) recruit the complex (C3-B), TtC3-TtC2/Bf-1, or TtC3-TtC2/Bf-2. The second C3 convertase (C3b-Bb), TtC3b-TtC2/Bf-1b or TtC3b-TtC2/Bf-2b complex, may be formed by an unidentified protease on the surfaces of Gram-positive bacteria and fungi. TL-1 (Tachylectin-1) and possibly TPL-1 (Tachypleus plasma lectin-1) also promote TtC3b deposition on the three types of microbes, especially on fungi.
Mentions: Figure 9 shows a schematic model of TtC3b deposition on the surfaces of three types of microbes. The Ca2+-dependent lectins (TtCRP-1 and TL-5A) and the Ca2+-independent lectin (TLP-1/TL-1) seem to recruit the complex between TtC3 and TtC2/Bf-1 or TtC2/Bf-2 (C3(H2O)B) on Gram-positive bacteria and fungi, respectively, to convert the complex to a TtC3 convertase (C3bBb). These lectins seem to work as carries for the C3bBb-like convertase to the surface of microbes with quite specific binding activities. However, these lectins have broad binding specificities against microbial cell wall substances and bind not only to Gram-positive bacteria but also to Gram-negative bacteria [7], [9], [10], [13], [14]. In addition, TPL-1 and TL-1 bind to saccharides, such as N-acetyl-glucosamine, galactose, and mannan [13], [14], and possibly bind to β-1,3-D-glucan, a major cell wall component of fungi. Therefore, the binding specificities of these lectins are not sufficient to explain the microbe specific TtC3b deposition. Unknown proteins may be additionally required to determine the specific target of these lectins.

Bottom Line: Complement C3 plays an essential role in the opsonization of pathogens in the mammalian complement system, whereas the molecular mechanism underlying C3 activation in invertebrates remains unknown.TtC2/Bf-1 and TtC2/Bf-2 were synthesized and glycosylated in hemocytes and secreted to hemolymph plasma, which existed in a complex with C3 (TtC3), and their activation by microbes was absolutely Mg(2+)-dependent.We conclude that plasma lectins and factor C play key roles in microbe-specific TtC3b deposition in a C2/factor B-dependent or -independent manner.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Systems Life Sciences, Kyushu University, Fukuoka, Japan.

ABSTRACT
Complement C3 plays an essential role in the opsonization of pathogens in the mammalian complement system, whereas the molecular mechanism underlying C3 activation in invertebrates remains unknown. To understand the molecular mechanism of C3b deposition on microbes, we characterized two types of C2/factor B homologs (designated TtC2/Bf-1 and TtC2/Bf-2) identified from the horseshoe crab Tachypleus tridentatus. Although the domain architectures of TtC2/Bf-1 and TtC2/Bf-2 were identical to those of mammalian homologs, they contained five-repeated and seven-repeated complement control protein domains at their N-terminal regions, respectively. TtC2/Bf-1 and TtC2/Bf-2 were synthesized and glycosylated in hemocytes and secreted to hemolymph plasma, which existed in a complex with C3 (TtC3), and their activation by microbes was absolutely Mg(2+)-dependent. Flow cytometric analysis revealed that TtC3b deposition was Mg(2+)-dependent on Gram-positive bacteria or fungi, but not on Gram-negative bacteria. Moreover, this analysis demonstrated that Ca(2+)-dependent lectins (C-reactive protein-1 and tachylectin-5A) were required for TtC3b deposition on Gram-positive bacteria, and that a Ca(2+)-independent lectin (Tachypleus plasma lectin-1) was definitely indispensable for TtC3b deposition on fungi. In contrast, a horseshoe crab lipopolysaccharide-sensitive protease factor C was necessary and sufficient to deposit TtC3b on Gram-negative bacteria. We conclude that plasma lectins and factor C play key roles in microbe-specific TtC3b deposition in a C2/factor B-dependent or -independent manner.

Show MeSH
Related in: MedlinePlus