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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]
+ Show Author Affiliations
  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.
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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. https://doi.org/10.1074/jbc.M117.802736
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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. https://doi.org/10.1074/jbc.M117.802736. https://www.osti.gov/servlets/purl/1438877.
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@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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https://doi.org/10.1074/jbc.M117.802736
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Works referenced in this record:

Apolipoprotein E Is the Major Physiological Activator of Lecithin−Cholesterol Acyltransferase (LCAT) on Apolipoprotein B Lipoproteins †
journal, January 2005

  • Zhao, Yue; Thorngate, Fayanne E.; Weisgraber, Karl H.
  • Biochemistry, Vol. 44, Issue 3
  • DOI: 10.1021/bi0481489

Surface Plasmon Resonance Biosensor Studies of Human Wild-Type and Mutant Lecithin Cholesterol Acyltransferase Interactions with Lipoproteins †
journal, November 1999

  • Jin, Lihua; Shieh, Jeng-Jong; Grabbe, Edith
  • Biochemistry, Vol. 38, Issue 47
  • DOI: 10.1021/bi9916729

Combined monogenic hypercholesterolemia and hypoalphalipoproteinemia caused by mutations in LDL-R and LCAT genes
journal, September 2005

Lecithin cholesterol acyltransferase
journal, December 2000

A solution for the best rotation to relate two sets of vectors
journal, September 1976

Overview of the CCP 4 suite and current developments
journal, March 2011

  • Winn, Martyn D.; Ballard, Charles C.; Cowtan, Kevin D.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
  • DOI: 10.1107/S0907444910045749

Investigating Monoclonal Antibody Aggregation Using a Combination of H/DX-MS and Other Biophysical Measurements
journal, December 2013

  • Iacob, Roxana E.; Bou-Assaf, George M.; Makowski, Lee
  • Journal of Pharmaceutical Sciences, Vol. 102, Issue 12
  • DOI: 10.1002/jps.23754

Molecular Genetic Study of Finns With Hypoalphalipoproteinemia and Hyperalphalipoproteinemia: A Novel Gly
journal, April 1998

  • Miettinen, H. E.; Gylling, H.; Tenhunen, J.
  • Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 18, Issue 4
  • DOI: 10.1161/01.ATV.18.4.591

Agonistic Human Antibodies Binding to Lecithin-Cholesterol Acyltransferase Modulate High Density Lipoprotein Metabolism
journal, December 2015

  • Gunawardane, Ruwanthi N.; Fordstrom, Preston; Piper, Derek E.
  • Journal of Biological Chemistry, Vol. 291, Issue 6
  • DOI: 10.1074/jbc.M115.672790

The open conformation of a Pseudomonas lipase
journal, February 1997

The high-resolution crystal structure of human LCAT
journal, July 2015

  • Piper, Derek E.; Romanow, William G.; Gunawardane, Ruwanthi N.
  • Journal of Lipid Research, Vol. 56, Issue 9
  • DOI: 10.1194/jlr.M059873

Distribution of lecithin-cholesterol acyltransferase (LCAT) in human plasma lipoprotein fractions. Evidence for the association of active LCAT with low density lipoproteins
journal, August 1982

  • Chen, Ching-Hong; Albers, John J.
  • Biochemical and Biophysical Research Communications, Vol. 107, Issue 3
  • DOI: 10.1016/0006-291X(82)90633-7

Differential phenotypic expression by three mutant alleles in familial lecithin:cholesterol acyltransferase deficiency
journal, September 1991

Structural basis of the chiral selectivity of Pseudomonas cepacia lipase
journal, June 1998

Structural and Functional Properties of Two Mutants of Lecithin-Cholesterol Acyltransferase (T123I and N228K)
journal, December 1998

  • Adimoolam, Shanthi; Jin, Lihua; Grabbe, Edith
  • Journal of Biological Chemistry, Vol. 273, Issue 49
  • DOI: 10.1074/jbc.273.49.32561

In Vitro Expression of Structural Defects in the Lecithin-Cholesterol Acyltransferase Gene
journal, April 1995

  • Klein, Hanns-Georg; Duverger, Nicolas; Albers, John J.
  • Journal of Biological Chemistry, Vol. 270, Issue 16
  • DOI: 10.1074/jbc.270.16.9443

Interfacial activation of the lipase–procolipase complex by mixed micelles revealed by X-ray crystallography
journal, April 1993

  • van Tilbeurgh, Herman; Egloff, Marie-Pierre; Martinez, Chrislaine
  • Nature, Vol. 362, Issue 6423
  • DOI: 10.1038/362814a0

Scaling and assessment of data quality
journal, December 2005

  • Evans, Philip
  • Acta Crystallographica Section D Biological Crystallography, Vol. 62, Issue 1, p. 72-82
  • DOI: 10.1107/S0907444905036693

Production and Characterization ofa Reconstituted High Density Lipoprotein for Therapeutic Applications
journal, September 1996

Crystal Structure of Pseudomonas aeruginosa Lipase in the Open Conformation : THE PROTOTYPE FOR FAMILY I.1 OF BACTERIAL LIPASES
journal, July 2000

  • Nardini, Marco; Lang, Dietmar A.; Liebeton, Klaus
  • Journal of Biological Chemistry, Vol. 275, Issue 40
  • DOI: 10.1074/jbc.M003903200

Genetic lecithin:cholesterol acyltransferase deficiency and cardiovascular disease
journal, June 2012

XDS
journal, January 2010

  • Kabsch, Wolfgang
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2
  • DOI: 10.1107/S0907444909047337

A unique genetic and biochemical presentation of fish-eye disease.
journal, December 1995

  • Kuivenhoven, J. A.; van Voorst tot Voorst, E. J.; Wiebusch, H.
  • Journal of Clinical Investigation, Vol. 96, Issue 6
  • DOI: 10.1172/JCI118348

Analysis of human lecithin–cholesterol acyltransferase activity by carboxyl-terminal truncation
journal, February 1997

  • Lee, Young-Phil; Adimoolam, Shanthi; Liu, Ming
  • Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, Vol. 1344, Issue 3
  • DOI: 10.1016/S0005-2760(96)00149-X

UCSF Chimera?A visualization system for exploratory research and analysis
journal, January 2004

  • Pettersen, Eric F.; Goddard, Thomas D.; Huang, Conrad C.
  • Journal of Computational Chemistry, Vol. 25, Issue 13
  • DOI: 10.1002/jcc.20084

A fluorescence method to detect and quantitate sterol esterification by lecithin:cholesterol acyltransferase
journal, October 2013

  • Homan, Reynold; Esmaeil, Nadia; Mendelsohn, Laurel
  • Analytical Biochemistry, Vol. 441, Issue 1
  • DOI: 10.1016/j.ab.2013.06.018

A protein cofactor of lecithin:Cholesterol acyltransferase
journal, February 1972

  • Fielding, C. J.; Shore, V. G.; Fielding, P. E.
  • Biochemical and Biophysical Research Communications, Vol. 46, Issue 4
  • DOI: 10.1016/0006-291X(72)90776-0

A molecular defect causing fish eye disease: an amino acid exchange in lecithin-cholesterol acyltransferase (LCAT) leads to the selective loss of alpha-LCAT activity.
journal, June 1991

  • Funke, H.; von Eckardstein, A.; Pritchard, P. H.
  • Proceedings of the National Academy of Sciences, Vol. 88, Issue 11
  • DOI: 10.1073/pnas.88.11.4855

Comparative Protein Modelling by Satisfaction of Spatial Restraints
journal, December 1993

High-Speed and High-Resolution UPLC Separation at Zero Degrees Celsius
journal, September 2008

  • Wales, Thomas E.; Fadgen, Keith E.; Gerhardt, Geoff C.
  • Analytical Chemistry, Vol. 80, Issue 17
  • DOI: 10.1021/ac8008862

Simultaneous Qualitative and Quantitative Analysis of the Escherichia coli Proteome : A Sweet Tale
journal, January 2006

  • Silva, Jeffrey C.; Denny, Richard; Dorschel, Craig
  • Molecular & Cellular Proteomics, Vol. 5, Issue 4
  • DOI: 10.1074/mcp.M500321-MCP200

Structural Differences Between Wild-type and Fish Eye Disease Mutant of Lecithin:cholesterol Acyltransferase
journal, August 2006

  • Reshetnyak, Yana; Tchedre, Kissaou T.; Nair, Maya P.
  • Journal of Biomolecular Structure and Dynamics, Vol. 24, Issue 1
  • DOI: 10.1080/07391102.2006.10507101

A model for interfacial activation in lipases from the structure of a fungal lipase-inhibitor complex
journal, June 1991

  • Brzozowski, A. M.; Derewenda, U.; Derewenda, Z. S.
  • Nature, Vol. 351, Issue 6326
  • DOI: 10.1038/351491a0

Hypoalphalipoproteinemia Resembling Fish Eye Disease
journal, January 1987

Deletion of N-terminal amino acids from human lecithin:cholesterol acyltransferase differentially affects enzyme activity toward α- and β-substrate lipoproteins
journal, March 2003

  • Vickaryous, Nicola K.; Teh, Evelyn M.; Stewart, Bruce
  • Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, Vol. 1646, Issue 1-2
  • DOI: 10.1016/S1570-9639(03)00005-0

MolProbity : all-atom structure validation for macromolecular crystallography
journal, December 2009

  • Chen, Vincent B.; Arendall, W. Bryan; Headd, Jeffrey J.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1
  • DOI: 10.1107/S0907444909042073

Lecithin Cholesterol Acyltransferase: An Anti- or Pro-atherogenic Factor?
journal, February 2011

  • Rousset, Xavier; Shamburek, Robert; Vaisman, Boris
  • Current Atherosclerosis Reports, Vol. 13, Issue 3
  • DOI: 10.1007/s11883-011-0171-6

Features and development of Coot
journal, March 2010

  • Emsley, P.; Lohkamp, B.; Scott, W. G.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
  • DOI: 10.1107/S0907444910007493

Molecular characterization of two patients with severe LCAT deficiency
journal, March 2007

  • Charlton-Menys, V.; Pisciotta, L.; Durrington, P. N.
  • Nephrology Dialysis Transplantation, Vol. 22, Issue 8
  • DOI: 10.1093/ndt/gfm311

REFMAC 5 for the refinement of macromolecular crystal structures
journal, March 2011

  • Murshudov, Garib N.; Skubák, Pavol; Lebedev, Andrey A.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
  • DOI: 10.1107/S0907444911001314

The Utility of Hydrogen/Deuterium Exchange Mass Spectrometry in Biopharmaceutical Comparability Studies
journal, June 2011

  • Houde, Damian; Berkowitz, Steven A.; Engen, John R.
  • Journal of Pharmaceutical Sciences, Vol. 100, Issue 6
  • DOI: 10.1002/jps.22432

Structure and function of lysosomal phospholipase A 2 : identification of the catalytic triad and the role of cysteine residues
journal, August 2005

Structure and function of lysosomal phospholipase A2 and lecithin:cholesterol acyltransferase
journal, March 2015

  • Glukhova, Alisa; Hinkovska-Galcheva, Vania; Kelly, Robert
  • Nature Communications, Vol. 6, Issue 1
  • DOI: 10.1038/ncomms7250

The detection, correlation, and comparison of peptide precursor and product ions from data independent LC-MS with data dependant LC-MS/MS
journal, March 2009

  • Geromanos, Scott J.; Vissers, Johannes P. C.; Silva, Jeffrey C.
  • PROTEOMICS, Vol. 9, Issue 6
  • DOI: 10.1002/pmic.200800562

Headgroup specificity of lecithin cholesterol acyltransferase for monomeric and vesicular phospholipids
journal, July 2000

  • Christiaens, Bart; Vanloo, Berlinda; Gouyette, Catherine
  • Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1486, Issue 2-3
  • DOI: 10.1016/S1388-1981(00)00075-5

The Influence of Adnectin Binding on the Extracellular Domain of Epidermal Growth Factor Receptor
journal, September 2014

  • Iacob, Roxana E.; Chen, Guodong; Ahn, Joomi
  • Journal of The American Society for Mass Spectrometry, Vol. 25, Issue 12
  • DOI: 10.1007/s13361-014-0973-1

Activation of lecithin:cholesterol acyltransferase by HDL ApoA-I central helices
journal, February 2009

  • Sorci-Thomas, Mary G.; Bhat, Shaila; Thomas, Michael J.
  • Clinical Lipidology, Vol. 4, Issue 1
  • DOI: 10.2217/17584299.4.1.113

Pepsin Immobilized on High-Strength Hybrid Particles for Continuous Flow Online Digestion at 10 000 psi
journal, August 2012

  • Ahn, Joomi; Jung, Moon Chul; Wyndham, Kevin
  • Analytical Chemistry, Vol. 84, Issue 16
  • DOI: 10.1021/ac301749h

Phaser crystallographic software
journal, July 2007

  • McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D.
  • Journal of Applied Crystallography, Vol. 40, Issue 4
  • DOI: 10.1107/S0021889807021206

Lecithin: cholesterol acyltransferase – from biochemistry to role in cardiovascular disease
journal, January 2009

  • Rousset, Xavier; Vaisman, Boris; Amar, Marcelo
  • Current Opinion in Endocrinology, Diabetes and Obesity, Vol. 16, Issue 2
  • DOI: 10.1097/MED.0b013e328329233b

The plasma lecithin:cholesterol acyltransferase reaction
journal, March 1968

The molecular pathology of lecithin:cholesterol acyltransferase (LCAT) deficiency syndromes
journal, February 1997

Effects of natural mutations in lecithin:cholesterol acyltransferase on the enzyme structure and activity
journal, January 1999

Relationship between structure and biochemical phenotype of lecithin:cholesterol acyltransferase (LCAT) mutants causing fish-eye disease
journal, May 2000

Effects of carboxy-terminal truncation on human lecithin:cholesterol acyltransferase activity.
journal, January 1996

In vitro expression of natural mutants of human lecithin:cholesterol acyltransferase.
journal, May 1995

Production and Characterization of a Reconstituted High Density Lipoprotein for Therapeutic Applications
journal, January 1996

  • Lerch, Peter G.; Förtsch, Vreni; Hodler, Gerhard
  • Vox Sanguinis, Vol. 71, Issue 3
  • DOI: 10.1159/000462047
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