Premium
High‐Fidelity Protein Targeting into Membrane Lipid Microdomains in Living Cells
Author(s) -
Beutel Oliver,
Nikolaus Jörg,
Birkholz Oliver,
You Changjiang,
Schmidt Thomas,
Herrmann Andreas,
Piehler Jacob
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201306328
Subject(s) - tris , chemistry , nitrilotriacetic acid , lipid microdomain , conjugated system , alkyl , vesicle , membrane , lipid bilayer , lysine , biochemistry , biophysics , organic chemistry , biology , amino acid , polymer , chelation
Lipid analogues carrying three nitrilotriacetic acid (tris‐NTA) head groups were developed for the selective targeting of His‐tagged proteins into liquid ordered (l o ) or liquid disordered (l d ) lipid phases. Strong partitioning into the l o phase of His‐tagged proteins bound to tris‐NTA conjugated to saturated alkyl chains (tris‐NTA DODA) was achieved, while tris‐NTA conjugated to an unsaturated alkyl chain (tris‐NTA SOA) predominantly resided in the l d phase. Interestingly, His‐tag‐mediated lipid crosslinking turned out to be required for efficient targeting into the l o phase by tris‐NTA DODA. Robust partitioning into l o phases was confirmed by using viral lipid mixtures and giant plasma membrane vesicles. Moreover, efficient protein targeting into l o and l d domains within the plasma membrane of living cells was demonstrated by single‐molecule tracking, thus establishing a highly generic approach for exploring lipid microdomains in situ.