2.3 Å resolution cryo-EM structure of human p97 and mechanism of allosteric inhibition

被引:270
作者
Banerjee, Soojay [1 ]
Bartesaghi, Alberto [1 ]
Merk, Alan [1 ]
Rao, Prashant [1 ]
Bulfer, Stacie L. [2 ]
Yan, Yongzhao [3 ]
Green, Neal [4 ]
Mroczkowski, Barbara [5 ]
Neitz, R. Jeffrey [2 ]
Wipf, Peter [3 ]
Falconieri, Veronica [1 ]
Deshaies, Raymond J. [6 ,7 ]
Milne, Jacqueline L. S. [1 ]
Huryn, Donna [3 ]
Arkin, Michelle [2 ]
Subramaniam, Sriram [1 ]
机构
[1] NCI, Cell Biol Lab, Bldg 37, Bethesda, MD 20892 USA
[2] Univ Calif San Francisco, Sch Pharm, Small Mol Discovery Ctr, Pharmaceut Chem, San Francisco, CA 94143 USA
[3] Univ Pittsburgh, Chem Divers Ctr, Pittsburgh, PA 15260 USA
[4] Leidos Biomed Res Inc, Frederick, MD 21702 USA
[5] NCI, Div Canc Treatment & Diag, Bethesda, MD 20892 USA
[6] CALTECH, Div Biol & Biol Engn, Pasadena, CA 91107 USA
[7] CALTECH, Howard Hughes Med Inst, Pasadena, CA 91107 USA
关键词
AAA-ATPASE; DOMAIN; ANALOGS; BINDING; P97/VCP; SYSTEM;
D O I
10.1126/science.aad7974
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
p97 is a hexameric AAA+ adenosine triphosphatase (ATPase) that is an attractive target for cancer drug development. We report cryo-electron microscopy (cryo-EM) structures for adenosine diphosphate (ADP)-bound, full-length, hexameric wild-type p97 in the presence and absence of an allosteric inhibitor at resolutions of 2.3 and 2.4 angstroms, respectively. We also report cryo-EM structures (at resolutions of similar to 3.3, 3.2, and 3.3 angstroms, respectively) for three distinct, coexisting functional states of p97 with occupancies of zero, one, or two molecules of adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S) per protomer. A large corkscrew-like change in molecular architecture, coupled with upward displacement of the N-terminal domain, is observed only when ATPgS is bound to both the D1 and D2 domains of the protomer. These cryo-EM structures establish the sequence of nucleotide-driven structural changes in p97 at atomic resolution. They also enable elucidation of the binding mode of an allosteric small-molecule inhibitor to p97 and illustrate how inhibitor binding at the interface between the D1 and D2 domains prevents propagation of the conformational changes necessary for p97 function.
引用
收藏
页码:871 / 875
页数:5
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