Structure of a specialized acyl carrier protein essential for lipid a biosynthesis with very long-chain fatty acids in open and closed conformations

Theresa A. Ramelot; Paolo Rossi; Farhad Forouhar; Hsiau Wei Lee; Yunhuang Yang; Shuisong Ni; Sarah Unser; Scott Lew; Jayaraman Seetharaman; Rong Xiao; Thomas B. Acton; John K. Everett; James H. Prestegard; John F. Hunt; Gaetano T. Montelione; Michael A. Kennedy

doi:10.1021/bi300546b

Structure of a specialized acyl carrier protein essential for lipid a biosynthesis with very long-chain fatty acids in open and closed conformations

Theresa A. Ramelot, Paolo Rossi, Farhad Forouhar, Hsiau Wei Lee, Yunhuang Yang, Shuisong Ni, Sarah Unser, Scott Lew, Jayaraman Seetharaman, Rong Xiao, Thomas B. Acton, John K. Everett, James H. Prestegard, John F. Hunt, Gaetano T. Montelione, Michael A. Kennedy

Rutgers, The State University

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

The solution nuclear magnetic resonance (NMR) structures and backbone ¹⁵N dynamics of the specialized acyl carrier protein (ACP), RpAcpXL, from Rhodopseudomonas palustris, in both the apo form and holo form modified by covalent attachment of 4′-phosphopantetheine at S37, are virtually identical, monomeric, and correspond to the closed conformation. The structures have an extra α-helix compared to the archetypical ACP from Escherichia coli, which has four helices, resulting in a larger opening to the hydrophobic cavity. Chemical shift differences between apo- and holo-RpAcpXL indicated some differences in the hinge region between α2 and α3 and in the hydrophobic cavity environment, but corresponding changes in nuclear Overhauser effect cross-peak patterns were not detected. In contrast to the NMR structures, apo-RpAcpXL was observed in an open conformation in crystals that diffracted to 2.0 Å resolution, which resulted from movement of α3. On the basis of the crystal structure, the predicted biological assembly is a homodimer. Although the possible biological significance of dimerization is unknown, there is potential that the resulting large shared hydrophobic cavity could accommodate the very long-chain fatty acid (28-30 carbons) that this specialized ACP is known to synthesize and transfer to lipid A. These structures are the first representatives of the AcpXL family and the first to indicate that dimerization may be important for the function of these specialized ACPs.

Original language	American English
Pages (from-to)	7239-7249
Number of pages	11
Journal	Biochemistry
Volume	51
Issue number	37
DOIs	https://doi.org/10.1021/bi300546b
State	Published - Sep 18 2012

ASJC Scopus subject areas

Biochemistry

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Ramelot, T. A., Rossi, P., Forouhar, F., Lee, H. W., Yang, Y., Ni, S., Unser, S., Lew, S., Seetharaman, J., Xiao, R., Acton, T. B., Everett, J. K., Prestegard, J. H., Hunt, J. F., Montelione, G. T., & Kennedy, M. A. (2012). Structure of a specialized acyl carrier protein essential for lipid a biosynthesis with very long-chain fatty acids in open and closed conformations. Biochemistry, 51(37), 7239-7249. https://doi.org/10.1021/bi300546b

@article{e0ed38bfa8c3415293bb19a608144869,

title = "Structure of a specialized acyl carrier protein essential for lipid a biosynthesis with very long-chain fatty acids in open and closed conformations",

abstract = "The solution nuclear magnetic resonance (NMR) structures and backbone 15N dynamics of the specialized acyl carrier protein (ACP), RpAcpXL, from Rhodopseudomonas palustris, in both the apo form and holo form modified by covalent attachment of 4′-phosphopantetheine at S37, are virtually identical, monomeric, and correspond to the closed conformation. The structures have an extra α-helix compared to the archetypical ACP from Escherichia coli, which has four helices, resulting in a larger opening to the hydrophobic cavity. Chemical shift differences between apo- and holo-RpAcpXL indicated some differences in the hinge region between α2 and α3 and in the hydrophobic cavity environment, but corresponding changes in nuclear Overhauser effect cross-peak patterns were not detected. In contrast to the NMR structures, apo-RpAcpXL was observed in an open conformation in crystals that diffracted to 2.0 {\AA} resolution, which resulted from movement of α3. On the basis of the crystal structure, the predicted biological assembly is a homodimer. Although the possible biological significance of dimerization is unknown, there is potential that the resulting large shared hydrophobic cavity could accommodate the very long-chain fatty acid (28-30 carbons) that this specialized ACP is known to synthesize and transfer to lipid A. These structures are the first representatives of the AcpXL family and the first to indicate that dimerization may be important for the function of these specialized ACPs.",

author = "Ramelot, {Theresa A.} and Paolo Rossi and Farhad Forouhar and Lee, {Hsiau Wei} and Yunhuang Yang and Shuisong Ni and Sarah Unser and Scott Lew and Jayaraman Seetharaman and Rong Xiao and Acton, {Thomas B.} and Everett, {John K.} and Prestegard, {James H.} and Hunt, {John F.} and Montelione, {Gaetano T.} and Kennedy, {Michael A.}",

year = "2012",

month = sep,

day = "18",

doi = "10.1021/bi300546b",

language = "American English",

volume = "51",

pages = "7239--7249",

journal = "Biochemistry",

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Ramelot, TA, Rossi, P, Forouhar, F, Lee, HW, Yang, Y, Ni, S, Unser, S, Lew, S, Seetharaman, J, Xiao, R, Acton, TB, Everett, JK, Prestegard, JH, Hunt, JF, Montelione, GT & Kennedy, MA 2012, 'Structure of a specialized acyl carrier protein essential for lipid a biosynthesis with very long-chain fatty acids in open and closed conformations', Biochemistry, vol. 51, no. 37, pp. 7239-7249. https://doi.org/10.1021/bi300546b

TY - JOUR

T1 - Structure of a specialized acyl carrier protein essential for lipid a biosynthesis with very long-chain fatty acids in open and closed conformations

AU - Ramelot, Theresa A.

AU - Rossi, Paolo

AU - Forouhar, Farhad

AU - Lee, Hsiau Wei

AU - Yang, Yunhuang

AU - Ni, Shuisong

AU - Unser, Sarah

AU - Lew, Scott

AU - Seetharaman, Jayaraman

AU - Xiao, Rong

AU - Acton, Thomas B.

AU - Everett, John K.

AU - Prestegard, James H.

AU - Hunt, John F.

AU - Montelione, Gaetano T.

AU - Kennedy, Michael A.

PY - 2012/9/18

Y1 - 2012/9/18

N2 - The solution nuclear magnetic resonance (NMR) structures and backbone 15N dynamics of the specialized acyl carrier protein (ACP), RpAcpXL, from Rhodopseudomonas palustris, in both the apo form and holo form modified by covalent attachment of 4′-phosphopantetheine at S37, are virtually identical, monomeric, and correspond to the closed conformation. The structures have an extra α-helix compared to the archetypical ACP from Escherichia coli, which has four helices, resulting in a larger opening to the hydrophobic cavity. Chemical shift differences between apo- and holo-RpAcpXL indicated some differences in the hinge region between α2 and α3 and in the hydrophobic cavity environment, but corresponding changes in nuclear Overhauser effect cross-peak patterns were not detected. In contrast to the NMR structures, apo-RpAcpXL was observed in an open conformation in crystals that diffracted to 2.0 Å resolution, which resulted from movement of α3. On the basis of the crystal structure, the predicted biological assembly is a homodimer. Although the possible biological significance of dimerization is unknown, there is potential that the resulting large shared hydrophobic cavity could accommodate the very long-chain fatty acid (28-30 carbons) that this specialized ACP is known to synthesize and transfer to lipid A. These structures are the first representatives of the AcpXL family and the first to indicate that dimerization may be important for the function of these specialized ACPs.

AB - The solution nuclear magnetic resonance (NMR) structures and backbone 15N dynamics of the specialized acyl carrier protein (ACP), RpAcpXL, from Rhodopseudomonas palustris, in both the apo form and holo form modified by covalent attachment of 4′-phosphopantetheine at S37, are virtually identical, monomeric, and correspond to the closed conformation. The structures have an extra α-helix compared to the archetypical ACP from Escherichia coli, which has four helices, resulting in a larger opening to the hydrophobic cavity. Chemical shift differences between apo- and holo-RpAcpXL indicated some differences in the hinge region between α2 and α3 and in the hydrophobic cavity environment, but corresponding changes in nuclear Overhauser effect cross-peak patterns were not detected. In contrast to the NMR structures, apo-RpAcpXL was observed in an open conformation in crystals that diffracted to 2.0 Å resolution, which resulted from movement of α3. On the basis of the crystal structure, the predicted biological assembly is a homodimer. Although the possible biological significance of dimerization is unknown, there is potential that the resulting large shared hydrophobic cavity could accommodate the very long-chain fatty acid (28-30 carbons) that this specialized ACP is known to synthesize and transfer to lipid A. These structures are the first representatives of the AcpXL family and the first to indicate that dimerization may be important for the function of these specialized ACPs.

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U2 - 10.1021/bi300546b

DO - 10.1021/bi300546b

M3 - Article

C2 - 22876860

SN - 0006-2960

VL - 51

SP - 7239

EP - 7249

JO - Biochemistry

JF - Biochemistry

IS - 37

ER -

Structure of a specialized acyl carrier protein essential for lipid a biosynthesis with very long-chain fatty acids in open and closed conformations

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