抄録
A 38 kDa β-1,3-glucanase allergen from Cryptomeria japonica pollen (CJP38) was recombinantly produced in E. coli and purified to homogeneity with the use of Ni-affinity resin. CJP38 hydrolyzed β-1,3-glucans such as CM-curdlan and laminarioligosaccharides in an endo-splitting manner. The optimum pH and temperature for β-1,3-glucanase activity were approximately 4.5 and 50 °C, respectively. The enzyme was stable at 30–60 °C and pH 4.0–10.5. Furthermore, CJP38 catalyzed a transglycosylation reaction to yield reaction products with a molecular weight higher than those of the starting laminarioligosaccharide substrates. The three-dimensional structure of CJP38 was determined using X-ray crystallography at 1.5 Å resolution. CJP38 exhibited the typical (β/α)8 TIM-barrel motif, similar to allergenic β-1,3-glucanases from banana (Mus a 5) and rubber tree latex (Hev b 2). Amino acid sequence alignment of these proteins indicated that the two-consensus IgE epitopes identified on the molecular surfaces of Mus a 5 and Hev b 2 were highly conserved in CJP38. Their conformations and surface locations were quite similar for these proteins. Sequence and structural conservation of these regions suggest that CJP38 is a candidate allergen responsible for the pollen-latex-fruit syndrome relating to Japanese cedar pollinosis.
| 元の言語 | 英語 |
|---|---|
| ページ（範囲） | 199-207 |
| ページ数 | 9 |
| ジャーナル | Molecular Immunology |
| 巻 | 116 |
| DOI | |
| 出版物ステータス | 出版済み - 12 2019 |
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All Science Journal Classification (ASJC) codes
- Immunology
- Molecular Biology
これを引用
Crystal structure and biochemical characterization of CJP38, a β-1,3-glucanase and allergen of Cryptomeria japonica pollen. / Takashima, Tomoya; Taku, Tomoki; Yamanaka, Tomoka; Fukamizo, Tamo; Numata, Tomoyuki; Ohnuma, Takayuki.
：: Molecular Immunology, 巻 116, 12.2019, p. 199-207.研究成果: ジャーナルへの寄稿 › 記事
}
TY - JOUR
T1 - Crystal structure and biochemical characterization of CJP38, a β-1,3-glucanase and allergen of Cryptomeria japonica pollen
AU - Takashima, Tomoya
AU - Taku, Tomoki
AU - Yamanaka, Tomoka
AU - Fukamizo, Tamo
AU - Numata, Tomoyuki
AU - Ohnuma, Takayuki
PY - 2019/12
Y1 - 2019/12
N2 - A 38 kDa β-1,3-glucanase allergen from Cryptomeria japonica pollen (CJP38) was recombinantly produced in E. coli and purified to homogeneity with the use of Ni-affinity resin. CJP38 hydrolyzed β-1,3-glucans such as CM-curdlan and laminarioligosaccharides in an endo-splitting manner. The optimum pH and temperature for β-1,3-glucanase activity were approximately 4.5 and 50 °C, respectively. The enzyme was stable at 30–60 °C and pH 4.0–10.5. Furthermore, CJP38 catalyzed a transglycosylation reaction to yield reaction products with a molecular weight higher than those of the starting laminarioligosaccharide substrates. The three-dimensional structure of CJP38 was determined using X-ray crystallography at 1.5 Å resolution. CJP38 exhibited the typical (β/α)8 TIM-barrel motif, similar to allergenic β-1,3-glucanases from banana (Mus a 5) and rubber tree latex (Hev b 2). Amino acid sequence alignment of these proteins indicated that the two-consensus IgE epitopes identified on the molecular surfaces of Mus a 5 and Hev b 2 were highly conserved in CJP38. Their conformations and surface locations were quite similar for these proteins. Sequence and structural conservation of these regions suggest that CJP38 is a candidate allergen responsible for the pollen-latex-fruit syndrome relating to Japanese cedar pollinosis.
AB - A 38 kDa β-1,3-glucanase allergen from Cryptomeria japonica pollen (CJP38) was recombinantly produced in E. coli and purified to homogeneity with the use of Ni-affinity resin. CJP38 hydrolyzed β-1,3-glucans such as CM-curdlan and laminarioligosaccharides in an endo-splitting manner. The optimum pH and temperature for β-1,3-glucanase activity were approximately 4.5 and 50 °C, respectively. The enzyme was stable at 30–60 °C and pH 4.0–10.5. Furthermore, CJP38 catalyzed a transglycosylation reaction to yield reaction products with a molecular weight higher than those of the starting laminarioligosaccharide substrates. The three-dimensional structure of CJP38 was determined using X-ray crystallography at 1.5 Å resolution. CJP38 exhibited the typical (β/α)8 TIM-barrel motif, similar to allergenic β-1,3-glucanases from banana (Mus a 5) and rubber tree latex (Hev b 2). Amino acid sequence alignment of these proteins indicated that the two-consensus IgE epitopes identified on the molecular surfaces of Mus a 5 and Hev b 2 were highly conserved in CJP38. Their conformations and surface locations were quite similar for these proteins. Sequence and structural conservation of these regions suggest that CJP38 is a candidate allergen responsible for the pollen-latex-fruit syndrome relating to Japanese cedar pollinosis.
UR - http://www.scopus.com/inward/record.url?scp=85074695462&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85074695462&partnerID=8YFLogxK
U2 - 10.1016/j.molimm.2019.10.016
DO - 10.1016/j.molimm.2019.10.016
M3 - Article
C2 - 31731097
AN - SCOPUS:85074695462
VL - 116
SP - 199
EP - 207
JO - Molecular Immunology
JF - Molecular Immunology
SN - 0161-5890
ER -