Substrate-assisted inhibition of ubiquitin-like protein-activating enzymes: the NEDD8 E1 inhibitor MLN4924 forms a NEDD8-AMP mimetic in situ.
The NEDD8-activating enzyme (NAE) initiates a protein homeostatic pathway essential for cancer cell growth and survival. MLN4924 is a selective inhibitor of NAE currently in clinical trials for the treatment of cancer. Here, we show that MLN4924 is a mechanism-based inhibitor of NAE and creates a covalent NEDD8-MLN4924 adduct catalyzed by the enzyme. The NEDD8-MLN4924 adduct resembles NEDD8 adenylate, the first intermediate in the NAE reaction cycle, but cannot be further utilized in subsequent intraenzyme reactions. The stability of the NEDD8-MLN4924 adduct within the NAE active site blocks enzyme activity, thereby accounting for the potent inhibition of the NEDD8 pathway by MLN4924. Importantly, we have determined that compounds resembling MLN4924 demonstrate the ability to form analogous adducts with other ubiquitin-like proteins (UBLs) catalyzed by their cognate-activating enzymes. These findings reveal insights into the mechanism of E1s and suggest a general strategy for selective inhibition of UBL conjugation pathways.
Brownell
James E
JE
Discovery, Millennium Pharmaceuticals, Inc., 40 Landsdowne Street, Cambridge, MA 02139, USA.
Sintchak
Michael D
MD
Gavin
James M
JM
Liao
Hua
H
Bruzzese
Frank J
FJ
Bump
Nancy J
NJ
Soucy
Teresa A
TA
Milhollen
Michael A
MA
Yang
Xiaofeng
X
Burkhardt
Anne L
AL
Ma
Jingya
J
Loke
Huay-Keng
HK
Lingaraj
Trupti
T
Wu
Dongyun
D
Hamman
Kristin B
KB
Spelman
James J
JJ
Cullis
Courtney A
CA
Langston
Steven P
SP
Vyskocil
Stepan
S
Sells
Todd B
TB
Mallender
William D
WD
Visiers
Irache
I
Li
Ping
P
Claiborne
Christopher F
CF
Rolfe
Mark
M
Bolen
Joseph B
JB
Dick
Lawrence R
LR
D000249:Adenosine Monophosphate / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D001667:Binding, Competitive; D045744:Cell Line, Tumor; D018360:Crystallography, X-Ray; D003517:Cyclopentanes / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D000075687:NEDD8 Protein; D017434:Protein Structure, Tertiary; D011743:Pyrimidines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D014452:Ubiquitins / Q000737:chemistry / Q000378:metabolism*
D003517:Cyclopentanes; D004791:Enzyme Inhibitors; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D011743:Pyrimidines; D014452:Ubiquitins; D000249:Adenosine Monophosphate; C539933:pevonedistat