Insight on the Molecular Envelope of Lipid-Bound Apolipoprotein E from Electron Paramagnetic Resonance Spectroscopy

Danny M. Hatters, John C Voss, Madhu S. Budamagunta, Yvonne N. Newhouse, Karl H. Weisgraber

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Although a high-resolution X-ray structure for the N-terminal domain of apolipoprotein E (apoE) in the lipid-free state has been solved, our knowledge of the structure of full-length apoE in a lipid-bound state is limited to an X-ray model fitting a molecular envelope at 10-Å resolution. To add molecular detail to the molecular envelope, we used cysteine mutagenesis to incorporate spin labels for analysis with electron paramagnetic resonance (EPR) spectroscopy. Twelve cysteine residues were introduced singly and in pairs at unique locations throughout apoE4 and labeled with an EPR spin probe. The labeled apoE4 was combined with dipalmitoylphosphatidylcholine, the particles were purified, and spectra were determined for 24 combinations (single and double) of the cysteine mutants. Data on the conformation, mobility, distance, and surface exposure of regions revealed by the cysteine probes were modeled into the molecular envelope of apoE bound to dipalmitoylphosphatidylcholine that had been determined by X-ray analysis. This EPR model of apoE in a native lipid-bound state validates the structural model derived from X-ray analysis and provides additional insight into apoE structure-function relationships.

Original languageEnglish (US)
Pages (from-to)261-271
Number of pages11
JournalJournal of Molecular Biology
Volume386
Issue number1
DOIs
StatePublished - Feb 13 2009

Fingerprint

Electron Spin Resonance Spectroscopy
Apolipoproteins E
Spectrum Analysis
Cysteine
Lipids
X-Rays
Apolipoprotein E4
1,2-Dipalmitoylphosphatidylcholine
Spin Labels
Structural Models
Mutagenesis

Keywords

  • lipoproteins
  • models
  • protein structure
  • spin labels

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Insight on the Molecular Envelope of Lipid-Bound Apolipoprotein E from Electron Paramagnetic Resonance Spectroscopy. / Hatters, Danny M.; Voss, John C; Budamagunta, Madhu S.; Newhouse, Yvonne N.; Weisgraber, Karl H.

In: Journal of Molecular Biology, Vol. 386, No. 1, 13.02.2009, p. 261-271.

Research output: Contribution to journalArticle

Hatters, Danny M. ; Voss, John C ; Budamagunta, Madhu S. ; Newhouse, Yvonne N. ; Weisgraber, Karl H. / Insight on the Molecular Envelope of Lipid-Bound Apolipoprotein E from Electron Paramagnetic Resonance Spectroscopy. In: Journal of Molecular Biology. 2009 ; Vol. 386, No. 1. pp. 261-271.
@article{a2dd8149c0784bb1b2916246e5b67ed7,
title = "Insight on the Molecular Envelope of Lipid-Bound Apolipoprotein E from Electron Paramagnetic Resonance Spectroscopy",
abstract = "Although a high-resolution X-ray structure for the N-terminal domain of apolipoprotein E (apoE) in the lipid-free state has been solved, our knowledge of the structure of full-length apoE in a lipid-bound state is limited to an X-ray model fitting a molecular envelope at 10-{\AA} resolution. To add molecular detail to the molecular envelope, we used cysteine mutagenesis to incorporate spin labels for analysis with electron paramagnetic resonance (EPR) spectroscopy. Twelve cysteine residues were introduced singly and in pairs at unique locations throughout apoE4 and labeled with an EPR spin probe. The labeled apoE4 was combined with dipalmitoylphosphatidylcholine, the particles were purified, and spectra were determined for 24 combinations (single and double) of the cysteine mutants. Data on the conformation, mobility, distance, and surface exposure of regions revealed by the cysteine probes were modeled into the molecular envelope of apoE bound to dipalmitoylphosphatidylcholine that had been determined by X-ray analysis. This EPR model of apoE in a native lipid-bound state validates the structural model derived from X-ray analysis and provides additional insight into apoE structure-function relationships.",
keywords = "lipoproteins, models, protein structure, spin labels",
author = "Hatters, {Danny M.} and Voss, {John C} and Budamagunta, {Madhu S.} and Newhouse, {Yvonne N.} and Weisgraber, {Karl H.}",
year = "2009",
month = "2",
day = "13",
doi = "10.1016/j.jmb.2008.12.040",
language = "English (US)",
volume = "386",
pages = "261--271",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Insight on the Molecular Envelope of Lipid-Bound Apolipoprotein E from Electron Paramagnetic Resonance Spectroscopy

AU - Hatters, Danny M.

AU - Voss, John C

AU - Budamagunta, Madhu S.

AU - Newhouse, Yvonne N.

AU - Weisgraber, Karl H.

PY - 2009/2/13

Y1 - 2009/2/13

N2 - Although a high-resolution X-ray structure for the N-terminal domain of apolipoprotein E (apoE) in the lipid-free state has been solved, our knowledge of the structure of full-length apoE in a lipid-bound state is limited to an X-ray model fitting a molecular envelope at 10-Å resolution. To add molecular detail to the molecular envelope, we used cysteine mutagenesis to incorporate spin labels for analysis with electron paramagnetic resonance (EPR) spectroscopy. Twelve cysteine residues were introduced singly and in pairs at unique locations throughout apoE4 and labeled with an EPR spin probe. The labeled apoE4 was combined with dipalmitoylphosphatidylcholine, the particles were purified, and spectra were determined for 24 combinations (single and double) of the cysteine mutants. Data on the conformation, mobility, distance, and surface exposure of regions revealed by the cysteine probes were modeled into the molecular envelope of apoE bound to dipalmitoylphosphatidylcholine that had been determined by X-ray analysis. This EPR model of apoE in a native lipid-bound state validates the structural model derived from X-ray analysis and provides additional insight into apoE structure-function relationships.

AB - Although a high-resolution X-ray structure for the N-terminal domain of apolipoprotein E (apoE) in the lipid-free state has been solved, our knowledge of the structure of full-length apoE in a lipid-bound state is limited to an X-ray model fitting a molecular envelope at 10-Å resolution. To add molecular detail to the molecular envelope, we used cysteine mutagenesis to incorporate spin labels for analysis with electron paramagnetic resonance (EPR) spectroscopy. Twelve cysteine residues were introduced singly and in pairs at unique locations throughout apoE4 and labeled with an EPR spin probe. The labeled apoE4 was combined with dipalmitoylphosphatidylcholine, the particles were purified, and spectra were determined for 24 combinations (single and double) of the cysteine mutants. Data on the conformation, mobility, distance, and surface exposure of regions revealed by the cysteine probes were modeled into the molecular envelope of apoE bound to dipalmitoylphosphatidylcholine that had been determined by X-ray analysis. This EPR model of apoE in a native lipid-bound state validates the structural model derived from X-ray analysis and provides additional insight into apoE structure-function relationships.

KW - lipoproteins

KW - models

KW - protein structure

KW - spin labels

UR - http://www.scopus.com/inward/record.url?scp=58549107197&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58549107197&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2008.12.040

DO - 10.1016/j.jmb.2008.12.040

M3 - Article

VL - 386

SP - 261

EP - 271

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 1

ER -