Crystal structures of human glycerol 3-phosphate dehydrogenase 1 (GPD1).
Homo sapiens L-alpha-glycerol-3-phosphate dehydrogenase 1 (GPD1) catalyzes the reversible biological conversion of dihydroxyacetone (DHAP) to glycerol-3-phosphate. The GPD1 protein was expressed in Escherichia coli, and purified as a fusion protein with glutathione S-transferase. Here we report the apoenzyme structure of GPD1 determined by multiwavelength anomalous diffraction phasing, and other complex structures with small molecules (NAD+ and DHAP) by the molecular replacement method. This enzyme structure is organized into two distinct domains, the N-terminal eight-stranded beta-sheet sandwich domain and the C-terminal helical substrate-binding domain. An electrophilic catalytic mechanism by the epsilon-NH3+ group of Lys204 is proposed on the basis of the structural analyses. In addition, the inhibitory effects of zinc and sulfate on GPDHs are assayed and discussed.
Ou
Xianjin
X
National Laboratory of Biomacromolecules, Institute of Biophysics (IBP), Chinese Academy of Sciences, Beijing 100101, China.
Ji
Chaoneng
C
Han
Xueqing
X
Zhao
Xiaodong
X
Li
Xuemei
X
Mao
Yumin
Y
Wong
Luet-Lok
LL
Bartlam
Mark
M
Rao
Zihe
Z
D002384:Catalysis; D003460:Crystallization; D018360:Crystallography, X-Ray; D004098:Dihydroxyacetone / Q000737:chemistry / Q000378:metabolism; D019281:Dimerization; D005993:Glycerolphosphate Dehydrogenase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D010710:Phosphates / Q000502:physiology; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D015032:Zinc / Q000502:physiology
D010710:Phosphates; D011994:Recombinant Proteins; D009243:NAD; D005993:Glycerolphosphate Dehydrogenase; D015032:Zinc; D004098:Dihydroxyacetone