Reconstituted IMPDH polymers accommodate both catalytically active and inactive conformations

被引:54
作者
Anthony, Sajitha A. [1 ]
Burrell, Anika L. [2 ]
Johnson, Matthew C. [2 ]
Duong-Ly, Krisna C. [1 ]
Kuo, Yin-Ming [1 ]
Simonet, Jacqueline C. [1 ]
Michener, Peter [3 ]
Andrews, Andrew [1 ]
Kollman, Justin M. [2 ]
Peterson, Jeffrey R. [1 ]
机构
[1] Fox Chase Canc Ctr, Canc Biol Program, Philadelphia, PA 19111 USA
[2] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
[3] Drexel Univ, Coll Med, Dept Biochem & Mol Biol, Philadelphia, PA 19102 USA
基金
美国国家卫生研究院;
关键词
INOSINE MONOPHOSPHATE DEHYDROGENASE; METABOLIC ENZYMES; SACCHAROMYCES-CEREVISIAE; ROD/RING STRUCTURES; MYCOPHENOLIC-ACID; REORGANIZATION; FILAMENTATION; PROTEINS; RINGS; YEAST;
D O I
10.1091/mbc.E17-04-0263
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Several metabolic enzymes undergo reversible polymerization into macromolecular assemblies. The function of these assemblies is often unclear, but in some cases they regulate enzyme activity and metabolic homeostasis. The guanine nucleotide biosynthetic enzyme inosine monophosphate dehydrogenase (IMPDH) forms octamers that polymerize into helical chains. In mammalian cells, IMPDH filaments can associate into micron-length assemblies. Polymerization and enzyme activity are regulated in part by binding of purine nucleotides to an allosteric regulatory domain. ATP promotes octamer polymerization, whereas guanosine triphosphate (GTP) promotes a compact, inactive conformation whose ability to polymerize is unknown. Also unclear is whether polymerization directly alters IMPDH catalytic activity. To address this, we identified point mutants of human IMPDH2 that either prevent or promote polymerization. Unexpectedly, we found that polymerized and nonassembled forms of recombinant IMPDH have comparable catalytic activity, substrate affinity, and GTP sensitivity and validated this finding in cells. Electron microscopy revealed that substrates and allosteric nucleotides shift the equilibrium between active and inactive conformations in both the octamer and the filament. Unlike other metabolic filaments, which selectively stabilize active or inactive conformations, recombinant IMPDH filaments accommodate multiple states. These conformational states are finely tuned by substrate availability and purine balance, while polymerization may allow cooperative transitions between states.
引用
收藏
页码:2600 / 2608
页数:9
相关论文
共 40 条
[1]   A Systematic Survey Identifies Prions and Illuminates Sequence Features of Prionogenic Proteins [J].
Alberti, Simon ;
Halfmann, Randal ;
King, Oliver ;
Kapila, Atul ;
Lindquist, Susan .
CELL, 2009, 137 (01) :146-158
[2]  
[Anonymous], 2011, THESIS ARTOIS
[3]   ConSurf 2016: an improved methodology to estimate and visualize evolutionary conservation in macromolecules [J].
Ashkenazy, Haim ;
Abadi, Shiran ;
Martz, Eric ;
Chay, Ofer ;
Mayrose, Itay ;
Pupko, Tal ;
Ben-Tal, Nir .
NUCLEIC ACIDS RESEARCH, 2016, 44 (W1) :W344-W350
[4]   Metabolic regulation via enzyme filamentation [J].
Aughey, Gabriel N. ;
Liu, Ji-Long .
CRITICAL REVIEWS IN BIOCHEMISTRY AND MOLECULAR BIOLOGY, 2016, 51 (04) :282-293
[5]   Large-scale filament formation inhibits the activity of CTP synthetase [J].
Barry, Rachael M. ;
Bitbol, Anne-Florence ;
Lorestani, Alexander ;
Charles, Emeric J. ;
Habrian, Chris H. ;
Hansen, Jesse M. ;
Li, Hsin-Jung ;
Baldwin, Enoch P. ;
Wingreen, Ned S. ;
Kollman, Justin M. ;
Gitai, Zemer .
ELIFE, 2014, 3 :1-19
[6]  
BEATY NB, 1983, J BIOL CHEM, V258, P3051
[7]   A nucleotide-controlled conformational switch modulates the activity of eukaryotic IMP dehydrogenases [J].
Buey, Ruben M. ;
Fernandez-Justel, David ;
Marcos-Alcalde, Inigo ;
Winter, Graeme ;
Gomez-Puertas, Paulino ;
Maria de Pereda, Jose ;
Luis Revuelta, Jose .
SCIENTIFIC REPORTS, 2017, 7
[8]   Guanine nucleotide binding to the Bateman domain mediates the allosteric inhibition of eukaryotic IMP dehydrogenases [J].
Buey, Ruben M. ;
Ledesma-Amaro, Rodrigo ;
Velazquez-campoy, Adrian ;
Balsera, Monica ;
Chagoyen, Monica ;
de Pereda, Jose M. ;
Revuelta, Jose L. .
NATURE COMMUNICATIONS, 2015, 6
[9]   'Rod and ring' formation from IMP dehydrogenase is regulated through the one-carbon metabolic pathway [J].
Calise, S. John ;
Purich, Daniel L. ;
Thuy Nguyen ;
Saleem, Dania A. ;
Krueger, Claire ;
Yin, Joyce D. ;
Chan, Edward K. L. .
JOURNAL OF CELL SCIENCE, 2016, 129 (15) :3042-3052
[10]   Glutamine deprivation initiates reversible assembly of mammalian rods and rings [J].
Calise, S. John ;
Carcamo, Wendy C. ;
Krueger, Claire ;
Yin, Joyce D. ;
Purich, Daniel L. ;
Chan, Edward K. L. .
CELLULAR AND MOLECULAR LIFE SCIENCES, 2014, 71 (15) :2963-2973