A Cell-Permeant Mimetic of NMN Activates SARM1 to Produce Cyclic ADP-Ribose and Induce Non-apoptotic Cell Death

Zhi Ying Zhao; Xu Jie Xie; Wan Hua Li; Jun Liu; Zhe Chen; Ben Zhang; Ting Li; Song Lu Li; Jun Gang Lu; Liangren Zhang; Li-He Zhang; Zhengshuang Xu; Hon Cheung Lee; Yong Juan Zhao

doi:10.1016/j.isci.2019.05.001

A Cell-Permeant Mimetic of NMN Activates SARM1 to Produce Cyclic ADP-Ribose and Induce Non-apoptotic Cell Death

iScience. 2019 May 31:15:452-466. doi: 10.1016/j.isci.2019.05.001. Epub 2019 May 4.

Authors

Zhi Ying Zhao¹, Xu Jie Xie¹, Wan Hua Li¹, Jun Liu¹, Zhe Chen², Ben Zhang¹, Ting Li¹, Song Lu Li¹, Jun Gang Lu³, Liangren Zhang², Li-He Zhang², Zhengshuang Xu¹, Hon Cheung Lee⁴, Yong Juan Zhao⁵

Affiliations

¹ State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055, China.
² State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
³ Agilent Technologies (China) Co.,Ltd, Guangzhou 510613, China.
⁴ State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055, China. Electronic address: leehoncheung@gmail.com.
⁵ State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055, China. Electronic address: zhaoyongjuan@pku.edu.cn.

Abstract

SARM1, an NAD-utilizing enzyme, regulates axonal degeneration. We show that CZ-48, a cell-permeant mimetic of NMN, activated SARM1 in vitro and in cellulo to cyclize NAD and produce a Ca²⁺ messenger, cADPR, with similar efficiency as NMN. Knockout of NMN-adenylyltransferase elevated cellular NMN and activated SARM1 to produce cADPR, confirming NMN was its endogenous activator. Determinants for the activating effects and cell permeability of CZ-48 were identified. CZ-48 activated SARM1 via a conformational change of the auto-inhibitory domain and dimerization of its catalytic domain. SARM1 catalysis was similar to CD38, despite having no sequence similarity. Both catalyzed similar set of reactions, but SARM1 had much higher NAD-cyclizing activity, making it more efficient in elevating cADPR. CZ-48 acted selectively, activating SARM1 but inhibiting CD38. In SARM1-overexpressing cells, CZ-48 elevated cADPR, depleted NAD and ATP, and induced non-apoptotic death. CZ-48 is a specific modulator of SARM1 functions in cells.

Keywords: Biochemical Mechanism; Biochemistry; Enzymology.