Structural insights into lipoprotein N-acylation by Escherichia coli apolipoprotein N-acyltransferase
Abstract
Gram-negative bacteria express a diverse array of lipoproteins that are essential for various aspects of cell growth and virulence, including nutrient uptake, signal transduction, adhesion, conjugation, sporulation, and outer membrane protein folding. Lipoprotein maturation requires the sequential activity of three enzymes that are embedded in the cytoplasmic membrane. First, phosphatidylglycerol:prolipoprotein diacylglyceryl transferase (Lgt) recognizes a conserved lipobox motif within the prolipoprotein signal sequence and catalyzes the addition of diacylglycerol to an invariant cysteine. The signal sequence is then cleaved by signal peptidase II (LspA) to give an N-terminal S-diacylglyceryl cysteine. Finally, apolipoprotein N-acyltransferase (Lnt) catalyzes the transfer of the sn-1-acyl chain of phosphatidylethanolamine to this N-terminal cysteine, generating a mature, triacylated lipoprotein. Although structural studies of Lgt and LspA have yielded significant mechanistic insights into this essential biosynthetic pathway, the structure of Lnt has remained elusive. Here, we present crystal structures of wild-type and an active-site mutant of Escherichia coli Lnt. The structures reveal a monomeric eight-transmembrane helix fold that supports a periplasmic carbon–nitrogen hydrolase domain containing a Cys–Glu–Lys catalytic triad. Two lipids are bound at the active site in the structures, and we propose a putative phosphate recognition site where a chloride ion is coordinated near the active site.more »
- Authors:
-
- Genentech Inc., San Francisco, CA (United States). Dept. of Structural Biology
- Univ. of California, Los Angeles, CA (United States)
- Genentech Inc., San Francisco, CA (United States). Dept. of Infectious Diseases
- Genentech Inc., San Francisco, CA (United States). Dept. of Biochemical and Cellular Pharmacology
- Genentech Inc., San Francisco, CA (United States). Dept. of Pathology
- Genentech Inc., San Francisco, CA (United States). Dept. of Translational Immunology
- Genentech Inc., San Francisco, CA (United States). Biomolecular Resource Group
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1437460
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America
- Additional Journal Information:
- Journal Volume: 114; Journal Issue: 30; Journal ID: ISSN 0027-8424
- Publisher:
- National Academy of Sciences
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; lipoprotein biosynthesis; apolipoprotein N-acyltransferase; crystal structure; enzyme mechanism
Citation Formats
Noland, Cameron L., Kattke, Michele D., Diao, Jingyu, Gloor, Susan L., Pantua, Homer, Reichelt, Mike, Katakam, Anand K., Yan, Donghong, Kang, Jing, Zilberleyb, Inna, Xu, Min, Kapadia, Sharookh B., and Murray, Jeremy M. Structural insights into lipoprotein N-acylation by Escherichia coli apolipoprotein N-acyltransferase. United States: N. p., 2017.
Web. doi:10.1073/pnas.1707813114.
Noland, Cameron L., Kattke, Michele D., Diao, Jingyu, Gloor, Susan L., Pantua, Homer, Reichelt, Mike, Katakam, Anand K., Yan, Donghong, Kang, Jing, Zilberleyb, Inna, Xu, Min, Kapadia, Sharookh B., & Murray, Jeremy M. Structural insights into lipoprotein N-acylation by Escherichia coli apolipoprotein N-acyltransferase. United States. https://doi.org/10.1073/pnas.1707813114
Noland, Cameron L., Kattke, Michele D., Diao, Jingyu, Gloor, Susan L., Pantua, Homer, Reichelt, Mike, Katakam, Anand K., Yan, Donghong, Kang, Jing, Zilberleyb, Inna, Xu, Min, Kapadia, Sharookh B., and Murray, Jeremy M. Tue .
"Structural insights into lipoprotein N-acylation by Escherichia coli apolipoprotein N-acyltransferase". United States. https://doi.org/10.1073/pnas.1707813114. https://www.osti.gov/servlets/purl/1437460.
@article{osti_1437460,
title = {Structural insights into lipoprotein N-acylation by Escherichia coli apolipoprotein N-acyltransferase},
author = {Noland, Cameron L. and Kattke, Michele D. and Diao, Jingyu and Gloor, Susan L. and Pantua, Homer and Reichelt, Mike and Katakam, Anand K. and Yan, Donghong and Kang, Jing and Zilberleyb, Inna and Xu, Min and Kapadia, Sharookh B. and Murray, Jeremy M.},
abstractNote = {Gram-negative bacteria express a diverse array of lipoproteins that are essential for various aspects of cell growth and virulence, including nutrient uptake, signal transduction, adhesion, conjugation, sporulation, and outer membrane protein folding. Lipoprotein maturation requires the sequential activity of three enzymes that are embedded in the cytoplasmic membrane. First, phosphatidylglycerol:prolipoprotein diacylglyceryl transferase (Lgt) recognizes a conserved lipobox motif within the prolipoprotein signal sequence and catalyzes the addition of diacylglycerol to an invariant cysteine. The signal sequence is then cleaved by signal peptidase II (LspA) to give an N-terminal S-diacylglyceryl cysteine. Finally, apolipoprotein N-acyltransferase (Lnt) catalyzes the transfer of the sn-1-acyl chain of phosphatidylethanolamine to this N-terminal cysteine, generating a mature, triacylated lipoprotein. Although structural studies of Lgt and LspA have yielded significant mechanistic insights into this essential biosynthetic pathway, the structure of Lnt has remained elusive. Here, we present crystal structures of wild-type and an active-site mutant of Escherichia coli Lnt. The structures reveal a monomeric eight-transmembrane helix fold that supports a periplasmic carbon–nitrogen hydrolase domain containing a Cys–Glu–Lys catalytic triad. Two lipids are bound at the active site in the structures, and we propose a putative phosphate recognition site where a chloride ion is coordinated near the active site. Based on these structures and complementary cell-based, biochemical, and molecular dynamics approaches, we propose a mechanism for substrate engagement and catalysis by E. coli Lnt.},
doi = {10.1073/pnas.1707813114},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 30,
volume = 114,
place = {United States},
year = {Tue Jul 11 00:00:00 EDT 2017},
month = {Tue Jul 11 00:00:00 EDT 2017}
}
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