Molecular insight into the potential functional role of pseudoenzyme GFOD1 via interaction with NKIRAS2  

作　　者：Jiawen Shi Xinyi Guo Chan Liu Yilun Wang Xiaobao Chen Guihua Wu Jianping Ding Tianlong Zhang 

机构地区：[1]Institute of Geriatrics,Affiliated Nantong Hospital of Shanghai University,Sixth People’s Hospital of Nantong,Shanghai Engineering Research Center of Organ Repair,School of Medicine,Shanghai University,Nantong 226011,China [2]State Key Laboratory of Molecular Biology,Shanghai Institute of Biochemistry and Cell Biology,Center for Excellence in Molecular Cell Science,Chinese Academy of Sciences,University of Chinese Academy of Sciences,Shanghai 200031,China [3]China-Japan Friendship Medical Research Institute,Shanghai University,Shanghai 200444,China

出　　处：《Acta Biochimica et Biophysica Sinica》2024年第9期1256-1266,共11页生物化学与生物物理学报（英文版）

基　　金：supported by the grants from the National Natural Science Foundation of China(Nos.32070766,32271247 and 32071190);the National Key Research and Development Program of China(Nos.2020YFA0509000 and 2020YFA0803200);the Jiangsu Commission of Health(No.LKZ2023018).

摘　　要：The glucose-fructose oxidoreductase/inositol dehydrogenase/rhizopine catabolism protein(Gfo/Idh/MocA)family includes a variety of oxidoreductases with a wide range of substrates that utilize NAD or NADP as redox cofactor.Human contains two members of this family,namely glucose-fructose oxidoreductase domain-containing protein 1 and 2(GFOD1 and GFOD2).While GFOD1 exhibits low tissue specificity,it is notably expressed in the brain,potentially linked to psychiatric disorders and severe diseases.Nevertheless,the specific function,cofactor preference,and enzymatic activity of GFOD1 remain largely unknown.In this work,we find that GFOD1 does not bind to either NAD or NADP.Crystal structure analysis unveils that GFOD1 exists as a typical homodimer resembling other family members,but lacks essential residues required for cofactor binding,suggesting that it may function as a pseudoenzyme.Exploration of GFOD1-interacting partners in proteomic database identifies NF-κB inhibitor-interacting Ras-like 2(NKIRAS2)as one potential candidate.Co-immunoprecipitation(co-IP)analysis indicates that GFOD1 interacts with both GTP-and GDP-bound forms of NKIRAS2.The predicted structural model of the GFOD1-NKIRAS2 complex is validated in cells using point mutants and shows that GFOD1 selectively recognizes the interswitch region of NKIRAS2.These findings reveal the distinct structural properties of GFOD1 and shed light on its potential functional role in cellular processes.

关 键 词：crystal structure glucose-fructose oxidoreductase GFoD1 glucose-fructose oxidoreductase/inositol dehydrogenase/rhizopine catabolism protein(Gfo/ldh/MocA)family NKIRAS2 pseudoenzyme 

分 类 号：Q55[生物学—生物化学]