Crystal and EM structures of human phosphoribosyl pyrophosphate synthase I (PRS1) provide novel insights into the disease-associated mutations

Peng Chen; Zheng Liu; Xuejuan Wang; Junhui Peng; Qianqian Sun; Jianzhong Li; Mingxing Wang; Liwen Niu; Zhiyong Zhang; Gang Cai; Maikun Teng; Xu Li

doi:10.1371/journal.pone.0120304

Crystal and EM structures of human phosphoribosyl pyrophosphate synthase I (PRS1) provide novel insights into the disease-associated mutations

PLoS One. 2015 Mar 17;10(3):e0120304. doi: 10.1371/journal.pone.0120304. eCollection 2015.

Authors

Peng Chen¹, Zheng Liu¹, Xuejuan Wang¹, Junhui Peng¹, Qianqian Sun¹, Jianzhong Li², Mingxing Wang¹, Liwen Niu¹, Zhiyong Zhang¹, Gang Cai¹, Maikun Teng¹, Xu Li¹

Affiliations

¹ Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230026, People's Republic of China; Key Laboratory of Structural Biology, Chinese Academy of Sciences, Hefei, Anhui 230026, People's Republic of China.
² Department of Otolaryngology Head and Neck Surgery, Fuzhou general hospital of Nanjing Command, PLA, Fuzhou 350025, China.

Abstract

Human PRS1, which is indispensable for the biosynthesis of nucleotides, deoxynucleotides and their derivatives, is associated directly with multiple human diseases because of single base mutation. However, a molecular understanding of the effect of these mutations is hampered by the lack of understanding of its catalytic mechanism. Here, we reconstruct the 3D EM structure of the PRS1 apo state. Together with the native stain EM structures of AMPNPP, AMPNPP and R5P, ADP and the apo states with distinct conformations, we suggest the hexamer is the enzymatically active form. Based on crystal structures, sequence analysis, mutagenesis, enzyme kinetics assays, and MD simulations, we reveal the conserved substrates binding motifs and make further analysis of all pathogenic mutants.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Crystallography
Humans
Molecular Dynamics Simulation
Molecular Sequence Data
Mutation*
Protein Binding
Ribose-Phosphate Pyrophosphokinase / chemistry*
Ribose-Phosphate Pyrophosphokinase / genetics
Ribose-Phosphate Pyrophosphokinase / metabolism
X-Rays

Substances

PRPS1 protein, human
Ribose-Phosphate Pyrophosphokinase

Grants and funding

Financial support for this project was provided by the Chinese Ministry of Science and Technology (Grant Nos. 2012CB917200), Chinese National Natural Science Foundation (Grant Nos. 31130018, 31370732, 31270014, 31270760 and U1432107), the Scientific Research Grant of Hefei Science Center of CAS (Grant No. 2015SRG-HSC042) and the Science and Technological Fund of Anhui Province for Outstanding Youth (Grant No. 1308085JGD08). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.