Molecular and biochemical characterizations of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis.
Fructose-1,6-bisphosphate aldolase (FbA) is a ubiquitous enzyme in glycolysis. In the present study, we screened out three distinct genes encoding FbA isozymes (CsFbAs, CsFbA-1/2/3) from Clonorchis sinensis (C. sinensis) and characterized their sequences and structures profiles as well as biochemical properties. The amino acid sequences of CsFbAs shared homology with those of Class I FbAs from other species. The putative quaternary structures revealed that CsFbA-2 and CsFbA-3 were tetramers, while CsFbA-1 was dimer. Recombinant CsFbA-2 and CsFbA-3 (rCsFbA-2/3) were confirmed to be Class I FbAs for their stable enzymatic activities in the presence of EDTA or metal ions. However, recombinant CsFbA-1 (rCsFbA-1) did not show the catalytic activity, which might be due to the inappropriate fold and interaction between its subunits. Both rCsFbA-2 and rCsFbA-3 showed similar enzymatic properties such as optimal temperatures and broad pH ranges that similar to human FbA isozymes. They showed relatively higher affinities for fructose-1,6-bisphosphate (FBP) than fructose-1-phosphate (F-1-P). Their kcat ratios of FBP to F-1-P were in accordance with those of human FbA-A or C. In addition, CsFbAs were differentially transcribed in the developmental stages of C. sinensis, suggesting their essential roles throughout the life stages. Extensive distribution of CsFbAs in adult worms indicated that ubiquitous activities of CsFbAs took place in these organs. Collectively, these results suggested that long-term parasitic environment might adapt these isozymes similar to host FbAs for metabolic requirement. Our study will provide new insight into CsFbAs in the glycometabolism of C. sinensis and relationship between the host and the parasite.
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Shan
S
Li
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Meng
M
Bian
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Xiaoyun
X
Wang
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Xueqing
X
Chen
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Zhizhi
Z
Xie
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Hengchang
H
Sun
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Feifei
F
Jia
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China; Department of Pathogen Biology, Hainan Medical College, Haikou, Hainan 571199, People's Republic of China.
Pei
P
Liang
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China; School of Nursing, Guangdong Medical College, Dongguan 523808, People's Republic of China.
Chenhui
C
Zhou
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Lei
L
He
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Qiang
Q
Mao
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Bo
B
Huang
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Chi
C
Liang
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Zhongdao
Z
Wu
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Xuerong
X
Li
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Jin
J
Xu
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China.
Yan
Y
Huang
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China; Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou 510080, People's Republic of China. Electronic address: yuhxteam@163.com.
Xinbing
X
Yu
D000818:Animals; D003004:Clonorchis sinensis / Q000201:enzymology* / Q000235:genetics; D004492:Edetic Acid / Q000378:metabolism; D020536:Enzyme Activators / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020869:Gene Expression Profiling; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008670:Metals / Q000378:metabolism; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D008670:Metals; D011994:Recombinant Proteins; D004492:Edetic Acid; D005634:Fructose-Bisphosphate Aldolase
Clonorchis sinensis; Enzymatic activity; Fructose-1,6-bisphosphate aldolase; Quaternary structure