EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Structural basis of the strict specificity of a bacterial GH31 α-1,3-glucosidase for nigerooligosaccharides.
ジャーナル・号・ページ	J Biol Chem, Vol. 298, Issue 5, Page 101827, Year 2022
掲載日	2022年3月12日
著者	Marina Ikegaya / Toshio Moriya / Naruhiko Adachi / Masato Kawasaki / Enoch Y Park / Takatsugu Miyazaki /
PubMed 要旨	Carbohydrate-active enzymes are involved in the degradation, biosynthesis, and modification of carbohydrates and vary with the diversity of carbohydrates. The glycoside hydrolase (GH) family 31 is ...Carbohydrate-active enzymes are involved in the degradation, biosynthesis, and modification of carbohydrates and vary with the diversity of carbohydrates. The glycoside hydrolase (GH) family 31 is one of the most diverse families of carbohydrate-active enzymes, containing various enzymes that act on α-glycosides. However, the function of some GH31 groups remains unknown, as their enzymatic activity is difficult to estimate due to the low amino acid sequence similarity between characterized and uncharacterized members. Here, we performed a phylogenetic analysis and discovered a protein cluster (GH31_u1) sharing low sequence similarity with the reported GH31 enzymes. Within this cluster, we showed that a GH31_u1 protein from Lactococcus lactis (LlGH31_u1) and its fungal homolog demonstrated hydrolytic activities against nigerose [α-D-Glcp-(1→3)-D-Glc]. The k/K values of LlGH31_u1 against kojibiose and maltose were 13% and 2.1% of that against nigerose, indicating that LlGH31_u1 has a higher specificity to the α-1,3 linkage of nigerose than other characterized GH31 enzymes, including eukaryotic enzymes. Furthermore, the three-dimensional structures of LlGH31_u1 determined using X-ray crystallography and cryogenic electron microscopy revealed that LlGH31_u1 forms a hexamer and has a C-terminal domain comprising four α-helices, suggesting that it contributes to hexamerization. Finally, crystal structures in complex with nigerooligosaccharides and kojibiose along with mutational analysis revealed the active site residues involved in substrate recognition in this enzyme. This study reports the first structure of a bacterial GH31 α-1,3-glucosidase and provides new insight into the substrate specificity of GH31 enzymes and the physiological functions of bacterial and fungal GH31_u1 members.
リンク	J Biol Chem / PubMed:35293315 / PubMed Central
手法	EM (単粒子) / X線回折
解像度	1.75 - 2.73 Å
構造データ	32571 7wlg EMDB-32571, PDB-7wlg: Cryo-EM structure of GH31 alpha-1,3-glucosidase from Lactococcus lactis subsp. cremoris 手法: EM (単粒子) / 解像度: 2.73 Å PDB-7wj9: Crystal structure of Lactococcus lactis subsp. cremoris GH31 alpha-1,3-glucosidase, P21 space group 手法: X-RAY DIFFRACTION / 解像度: 1.75 Å PDB-7wja: Crystal structure of Lactococcus lactis subsp. cremoris GH31 alpha-1,3-glucosidase, P6322 space group 手法: X-RAY DIFFRACTION / 解像度: 1.85 Å PDB-7wjb: Crystal structure of Lactococcus lactis subsp. cremoris GH31 alpha-1,3-glucosidase in complex with glucose 手法: X-RAY DIFFRACTION / 解像度: 2 Å PDB-7wjc: Crystal structure of Lactococcus lactis subsp. cremoris GH31 alpha-1,3-glucosidase mutant D394A in complex with nigerose 手法: X-RAY DIFFRACTION / 解像度: 1.75 Å PDB-7wjd: Crystal structure of Lactococcus lactis subsp. cremoris GH31 alpha-1,3-glucosidase mutant D394A in complex with nigerotriose 手法: X-RAY DIFFRACTION / 解像度: 1.8 Å PDB-7wje: Crystal structure of Lactococcus lactis subsp. cremoris GH31 alpha-1,3-glucosidase mutant D394A in complex with nigerotetraose 手法: X-RAY DIFFRACTION / 解像度: 1.8 Å PDB-7wjf: Crystal structure of Lactococcus lactis subsp. cremoris GH31 alpha-1,3-glucosidase mutant D394A in complex with kojibiose 手法: X-RAY DIFFRACTION / 解像度: 1.8 Å
化合物	ChemComp-XYL: Xylitol / キシリト-ル ChemComp-EDO: 1,2-ETHANEDIOL / エチレングリコ-ル ChemComp-HOH: WATER ChemComp-GLC: alpha-D-glucopyranose / α-D-グルコピラノ-ス ChemComp-BGC: beta-D-glucopyranose / β-D-グルコピラノ-ス
由来	lactococcus lactis subsp. cremoris mg1363 (乳酸菌)
キーワード	HYDROLASE / nigerose / glucose / glycoside hydrolase / GH31 / carbohydrate / TIM-barrel / hexamer / kojibiose