Bacterial β-Glucosidase Reveals the Structural and Functional Basis of Genetic Defects in Human Glucocerebrosidase 2 (GBA2).
Human glucosylcerebrosidase 2 (GBA2) of the CAZy family GH116 is responsible for the breakdown of glycosphingolipids on the cytoplasmic face of the endoplasmic reticulum and Golgi apparatus. Genetic defects in GBA2 result in spastic paraplegia and cerebellar ataxia, while cross-talk between GBA2 and GBA1 glucosylceramidases may affect Gaucher disease. Here, we report the first three-dimensional structure for any GH116 enzyme, Thermoanaerobacterium xylanolyticum TxGH116 β-glucosidase, alone and in complex with diverse ligands. These structures allow identification of the glucoside binding and active site residues, which are shown to be conserved with GBA2. Mutagenic analysis of TxGH116 and structural modeling of GBA2 provide a detailed structural and functional rationale for pathogenic missense mutations of GBA2.
Institute for Protein Research, Osaka University , 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.|School of Biochemistry, Institute of Science, Suranaree Univerity of Technology , Nakhon Ratchasima 30000, Thailand.|Center for Biomolecular Structure, Function and Application, Suranaree University of Technology , Nakhon Ratchasima 30000, Thailand.
Ratana
R
Charoenwattanasatien
School of Biochemistry, Institute of Science, Suranaree Univerity of Technology , Nakhon Ratchasima 30000, Thailand.|Center for Biomolecular Structure, Function and Application, Suranaree University of Technology , Nakhon Ratchasima 30000, Thailand.
Salila
S
Pengthaisong
Structural Biology Laboratory, Department of Chemistry, The University of York , York YO10 5DD, United Kingdom.
Imogen
I
Breen
Institute for Protein Research, Osaka University , 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Risa
R
Mutoh
Department of Chemistry, Faculty of Science, Rajamangala University of Technology , Thanyaburi, Pathum Thani 12110, Thailand.
Sompong
S
Sansenya
Center for Biomolecular Structure, Function and Application, Suranaree University of Technology , Nakhon Ratchasima 30000, Thailand.|Center for Scientific and Technological Equipment, Suranaree University of Technology , Nakhon Ratchasima 30000, Thailand.
Yanling
Y
Hua
Department of Biotechnology, Faculty of Agro-Industrial Technology, Kalasin University , Kalasin 46000, Thailand.
Anupong
A
Tankrathok
Structural Biology Laboratory, Department of Chemistry, The University of York , York YO10 5DD, United Kingdom.
Liang
L
Wu
Center for Biomolecular Structure, Function and Application, Suranaree University of Technology , Nakhon Ratchasima 30000, Thailand.|Synchrotron Light Research Institute , Nakhon Ratchasima 30000, Thailand.
Chomphunuch
C
Songsiriritthigul
Institute for Protein Research, Osaka University , 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Hideaki
H
Tanaka
School of Chemistry and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne , Parkville, Victoria 3010, Australia.
Spencer J
SJ
Williams
Structural Biology Laboratory, Department of Chemistry, The University of York , York YO10 5DD, United Kingdom.
Gideon J
GJ
Davies
Institute for Protein Research, Osaka University , 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Genji
G
Kurisu
School of Biochemistry, Institute of Science, Suranaree Univerity of Technology , Nakhon Ratchasima 30000, Thailand.|Center for Biomolecular Structure, Function and Application, Suranaree University of Technology , Nakhon Ratchasima 30000, Thailand.|Laboratory of Biochemistry, Chulabhorn Research Institute , Bangkok 10210, Thailand.
James R Ketudat
JR
Cairns
D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D005962:Glucosylceramidase; D006801:Humans; D020125:Mutation, Missense; D046950:Thermoanaerobacterium / Q000201:enzymology*; D001617:beta-Glucosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D001617:beta-Glucosidase; C517537:GBA2 protein, human; D005962:Glucosylceramidase