Cryo-EM structures of NPC1L1 reveal mechanisms of cholesterol transport and ezetimibe inhibition

Huang, Ching-Shin; Yu, Xinchao; Fordstrom, Preston; Choi, Kaylee; Chung, Ben C.; Roh, Soung-Hun; Chiu, Wah; Zhou, Mingyue; Min, Xiaoshan; Wang, Zhulun

doi:10.1126/sciadv.abb1989

Cryo-EM structures of NPC1L1 reveal mechanisms of cholesterol transport and ezetimibe inhibition

Journal Article · Fri Jun 19 00:00:00 EDT 2020 · Science Advances

DOI:https://doi.org/10.1126/sciadv.abb1989· OSTI ID:1647629

^[1]; ^[1]; ^[2]; ^[1]; Chung, Ben C. ^[1]; Roh, Soung-Hun ^[3]; Chiu, Wah ^[4]; Zhou, Mingyue ^[2]; ^[1]; ^[1]

Amgen Inc., South San Francisco, CA (United States). Dept. of Therapeutic Discovery
Amgen Inc., South San Francisco, CA (United States). Dept. of Cardiometabolic Disorders
Stanford Univ., CA (United States). Dept. of Bioengineering and James H. Clark Center
Stanford Univ., CA (United States). Dept. of Bioengineering and James H. Clark Center; SLAC National Accelerator Lab., Menlo Park, CA (United States). Division of Cryo-EM and Bioimaging

The intestinal absorption of cholesterol is mediated by a multipass membrane protein, Niemann-Pick C1-Like 1 (NPC1L1), the molecular target of a cholesterol lowering therapy ezetimibe. While ezetimibe gained Food and Drug Administration approval in 2002, its mechanism of action has remained unclear. Here, we present two cryo–electron microscopy structures of NPC1L1, one in its apo form and the other complexed with ezetimibe. The apo form represents an open state in which the N-terminal domain (NTD) interacts loosely with the rest of NPC1L1, leaving the NTD central cavity accessible for cholesterol loading. The ezetimibe-bound form signifies a closed state in which the NTD rotates ~60°, creating a continuous tunnel enabling cholesterol movement into the plasma membrane. Ezetimibe blocks cholesterol transport by occluding the tunnel instead of competing with cholesterol binding. These findings provide insight into the molecular mechanisms of NPC1L1-mediated cholesterol transport and ezetimibe inhibition, paving the way for more effective therapeutic development.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); National Research Foundation of Korea (NRF); USDOE Office of Science (SC)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1647629

Journal Information:: Science Advances, Journal Name: Science Advances Journal Issue: 25 Vol. 6; ISSN 2375-2548

Publisher:: AAASCopyright Statement

Country of Publication:: United States

Language:: English

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Additional file 1 of Dietary diosgenin transcriptionally down-regulated intestinal NPC1L1 expression to prevent cholesterol gallstone formation in mice Shen, Weiyi; Shao, Wentao; Wang, Qihan figshare https://doi.org/10.6084/m9.figshare.23590365.v1	image	January 2023

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