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Title: A retractable lid in lecithin:cholesterol acyltransferase provides a structural mechanism for activation by apolipoprotein A-I

Abstract

Lecithin:cholesterol acyltransferase (LCAT) plays a key role in reverse cholesterol transport by transferring an acyl group from phosphatidylcholine to cholesterol, promoting the maturation of high-density lipoproteins (HDL) from discoidal to spherical particles. LCAT is activated through an unknown mechanism by apolipoprotein A-I (apoA-I) and other mimetic peptides that form a belt around HDL. Here, we report the crystal structure of LCAT with an extended lid that blocks access to the active site, consistent with an inactive conformation. Residues Thr-123 and Phe-382 in the catalytic domain form a latch-like interaction with hydrophobic residues in the lid. Because these residues are mutated in genetic disease, lid displacement was hypothesized to be an important feature of apoA-I activation. Functional studies of site-directed mutants revealed that loss of latch interactions or the entire lid enhanced activity against soluble ester substrates, and hydrogen–deuterium exchange (HDX) mass spectrometry revealed that the LCAT lid is extremely dynamic in solution. Upon addition of a covalent inhibitor that mimics one of the reaction intermediates, there is an overall decrease in HDX in the lid and adjacent regions of the protein, consistent with ordering. As a result, these data suggest a model wherein the active site of LCAT is shieldedmore » from soluble substrates by a dynamic lid until it interacts with HDL to allow transesterification to proceed.« less

Authors:
 [1];  [2];  [3];  [1];  [2];  [1];  [1];  [1];  [2];  [1];  [4]
  1. Univ. of Michigan, Ann Arbor, MI (United States)
  2. MedImmune, Gaithersburg, MD (United States)
  3. Univ. of Michigan, Ann Arbor, MI (United States); Monash Univ., Parkville, VIC (Australia)
  4. Univ. of Michigan, Ann Arbor, MI (United States); Purdue Univ., West Lafayette, IN (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Inst. of Health; American Heart Assoc. Scientist Development Grant; American Heart Assoc. Postdoctoral Fellowship; Ruth L. Kirschstein NRSA
OSTI Identifier:
1438877
Grant/Contract Number:  
[HL071818; HL122416; AR056991; 13SDG17230049; 16POST27760002; 15POST24870001; F32HL131288]
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
[ Journal Volume: 292; Journal Issue: 49]; Journal ID: ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular Biology
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; acyltransferase; apolipoprotein; cholesterol; conformational change; crystallography; high-density lipoprotein (HDL); inhibitor; structural biology; HDX MS; lecithin

Citation Formats

Manthei, Kelly A., Ahn, Joomi, Glukhova, Alisa, Yuan, Wenmin, Larkin, Christopher, Manett, Taylor D., Chang, Louise, Shayman, James A., Axley, Milton J., Schwendeman, Anna, and Tesmer, John J. G. A retractable lid in lecithin:cholesterol acyltransferase provides a structural mechanism for activation by apolipoprotein A-I. United States: N. p., 2017. Web. doi:10.1074/jbc.M117.802736.
Manthei, Kelly A., Ahn, Joomi, Glukhova, Alisa, Yuan, Wenmin, Larkin, Christopher, Manett, Taylor D., Chang, Louise, Shayman, James A., Axley, Milton J., Schwendeman, Anna, & Tesmer, John J. G. A retractable lid in lecithin:cholesterol acyltransferase provides a structural mechanism for activation by apolipoprotein A-I. United States. doi:10.1074/jbc.M117.802736.
Manthei, Kelly A., Ahn, Joomi, Glukhova, Alisa, Yuan, Wenmin, Larkin, Christopher, Manett, Taylor D., Chang, Louise, Shayman, James A., Axley, Milton J., Schwendeman, Anna, and Tesmer, John J. G. Fri . "A retractable lid in lecithin:cholesterol acyltransferase provides a structural mechanism for activation by apolipoprotein A-I". United States. doi:10.1074/jbc.M117.802736. https://www.osti.gov/servlets/purl/1438877.
@article{osti_1438877,
title = {A retractable lid in lecithin:cholesterol acyltransferase provides a structural mechanism for activation by apolipoprotein A-I},
author = {Manthei, Kelly A. and Ahn, Joomi and Glukhova, Alisa and Yuan, Wenmin and Larkin, Christopher and Manett, Taylor D. and Chang, Louise and Shayman, James A. and Axley, Milton J. and Schwendeman, Anna and Tesmer, John J. G.},
abstractNote = {Lecithin:cholesterol acyltransferase (LCAT) plays a key role in reverse cholesterol transport by transferring an acyl group from phosphatidylcholine to cholesterol, promoting the maturation of high-density lipoproteins (HDL) from discoidal to spherical particles. LCAT is activated through an unknown mechanism by apolipoprotein A-I (apoA-I) and other mimetic peptides that form a belt around HDL. Here, we report the crystal structure of LCAT with an extended lid that blocks access to the active site, consistent with an inactive conformation. Residues Thr-123 and Phe-382 in the catalytic domain form a latch-like interaction with hydrophobic residues in the lid. Because these residues are mutated in genetic disease, lid displacement was hypothesized to be an important feature of apoA-I activation. Functional studies of site-directed mutants revealed that loss of latch interactions or the entire lid enhanced activity against soluble ester substrates, and hydrogen–deuterium exchange (HDX) mass spectrometry revealed that the LCAT lid is extremely dynamic in solution. Upon addition of a covalent inhibitor that mimics one of the reaction intermediates, there is an overall decrease in HDX in the lid and adjacent regions of the protein, consistent with ordering. As a result, these data suggest a model wherein the active site of LCAT is shielded from soluble substrates by a dynamic lid until it interacts with HDL to allow transesterification to proceed.},
doi = {10.1074/jbc.M117.802736},
journal = {Journal of Biological Chemistry},
number = [49],
volume = [292],
place = {United States},
year = {2017},
month = {10}
}

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    Works referencing / citing this record:

    Identification and functional analysis of missense mutations in the lecithin cholesterol acyltransferase gene in a Chilean patient with hypoalphalipoproteinemia
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