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PMID: 23931318 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Structure and topology of the huntingtin 1-17 membrane anchor by a combined solution and solid-state NMR approach.

Biophysical journal ·Vol. 105 ·No. 3 ·2013-08-06 ·Pages 699-710

Michalek M, Salnikov ES, Bechinger B

Abstract

The very amino-terminal domain of the huntingtin protein is directly located upstream of the protein's polyglutamine tract, plays a decisive role in several important properties of this large protein and in the development of Huntington's disease. This huntingtin 1-17 domain is on the one hand known to markedly increase polyglutamine aggregation rates and on the other hand has been shown to be involved in cellular membrane interactions. Here, we determined the high-resolution structure of huntingtin 1-17 in dodecyl phosphocholine micelles and the topology of its helical domain in oriented phosphatidylcholine bilayers. Using two-dimensional solution NMR spectroscopy the low-energy conformations of the polypeptide were identified in the presence of dodecyl phosphocholine detergent micelles. In a next step a set of four solid-state NMR angular restraints was obtained from huntingtin 1-17 labeled with (15)N and (2)H at selected sites. Of the micellar ensemble of helical conformations only a limited set agrees in quantitative detail with the solid-state angular restraints of huntingtin 1-17 obtained in supported planar lipid bilayers. Thereby, the solid-state NMR data were used to further refine the domain structure in phospholipid bilayers. At the same time its membrane topology was determined and different motional regimes of this membrane-associated domain were explored. The pronounced structural transitions of huntingtin 1-17 upon membrane-association result in a α-helical conformation from K6 to F17, i.e., up to the very start of the polyglutamine tract. This amphipathic helix is aligned nearly parallel to the membrane surface (tilt angle ∼77°) and is characterized by a hydrophobic ridge on one side and an alternation of cationic and anionic residues that run along the hydrophilic face of the helix. This arrangement facilitates electrostatic interactions between huntingtin 1-17 domains and possibly with the proximal polyglutamine tract.

Keywords
1 1 1 3 3 3-hexafluoro-2-propanol 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine CD DPC HFIP NES NOESY POPC SDS TFA TFE TOCSY circular dichroism dodecyl phosphocholine nuclear Overhauser effect spectroscopy nuclear export sequence sodium dodecylsulfate total correlation spectroscopy trifluoroacetic acid trifluoroethanol
MeSH Terms
Amino Acid Sequence Animals Humans Lipid Bilayers/chemistry Magnetic Resonance Spectroscopy Micelles Molecular Sequence Data Nerve Tissue Proteins/chemistry Protein Structure, Tertiary
Chemicals
Lipid Bilayers Micelles Nerve Tissue Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Michalek Matthias
Université de Strasbourg/CNRS, UMR, Institut de Chimie, France.
Salnikov Evgeniy S
Bechinger Burkhard
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
1542-0086
DOI
10.1016/j.bpj.2013.06.030
Published
2013-08-06
Pages
699-710
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC3736738
Subset
IM
External Links
PubMed source DOI: 10.1016/j.bpj.2013.06.030 PMC Full Text View journal details
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