Crystal structure and biochemical characterization of CJP38, a β-1,3-glucanase and allergen of Cryptomeria japonica pollen

Tomoya Takashima, Tomoki Taku, Tomoka Yamanaka, Tamo Fukamizo, Tomoyuki Numata, Takayuki Ohnuma

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)199-207
Number of pages9
JournalMolecular Immunology
Volume116
DOIs
Publication statusPublished - Dec 2019

Fingerprint

Cryptomeria
Latex
Pollen
Allergens
Hevea
Musa
Seasonal Allergic Rhinitis
Glucans
Sequence Alignment
X Ray Crystallography
Immunoglobulin E
Epitopes
Amino Acid Sequence
Fruit
Proteins
Molecular Weight
Escherichia coli
Temperature
Enzymes
carboxymethylcurdlan

All Science Journal Classification (ASJC) codes

  • Immunology
  • Molecular Biology

Cite this

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.

In: Molecular Immunology, Vol. 116, 12.2019, p. 199-207.

Research output: Contribution to journalArticle

Takashima, Tomoya ; Taku, Tomoki ; Yamanaka, Tomoka ; Fukamizo, Tamo ; Numata, Tomoyuki ; Ohnuma, Takayuki. / Crystal structure and biochemical characterization of CJP38, a β-1,3-glucanase and allergen of Cryptomeria japonica pollen. In: Molecular Immunology. 2019 ; Vol. 116. pp. 199-207.
@article{9198b88d2a6e4c6bb6b0e0f6dd1ecc03,
title = "Crystal structure and biochemical characterization of CJP38, a β-1,3-glucanase and allergen of Cryptomeria japonica pollen",
abstract = "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 {\AA} 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.",
author = "Tomoya Takashima and Tomoki Taku and Tomoka Yamanaka and Tamo Fukamizo and Tomoyuki Numata and Takayuki Ohnuma",
year = "2019",
month = "12",
doi = "10.1016/j.molimm.2019.10.016",
language = "English",
volume = "116",
pages = "199--207",
journal = "Molecular Immunology",
issn = "0161-5890",
publisher = "Elsevier Limited",

}

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 -