Azide- and diazirine-modified membrane lipids: Physicochemistry and applicability to study peptide/lipid interactions via cross-linking/mass spectrometry

被引:2
|
作者
Korn, Patricia [1 ]
Schwieger, Christian [2 ]
Gruhle, Kai [1 ,3 ]
Garamus, Vasil M. [4 ]
Meister, Annette [5 ,6 ]
Ihling, Christian [1 ,7 ]
Drescher, Simon [3 ,8 ]
机构
[1] Martin Luther Univ MLU Halle Wittenberg, Charles Tanford Prot Ctr, Inst Pharm Pharmaceut Chem & Bioanalyt, Kurt Mothes Str 3a, D-06120 Halle, Saale, Germany
[2] MLU Halle Wittenberg, Inst Chem, Danckelmann Pl 4, D-06120 Halle, Saale, Germany
[3] MLU Halle Wittenberg, Inst PharmacyBiophys Pharm, Wolfgang Langenbeck Str 4, D-06120 Halle, Saale, Germany
[4] Helmholtz Zentrum Hereon, Max Planck Str 1, D-21502 Geesthacht, Germany
[5] MLU Halle Wittenberg, Charles Tanford Prot Ctr, Interdisciplinary Res Ctr HALOmem, Kurt Mothes Str 3a, D-06120 Halle, Saale, Germany
[6] MLU Halle Wittenberg, Inst Biochem & Biotechnol Phys Biotechnol, Charles Tanford Prot Ctr, Kurt Mothes Str 3a, D-06120 Halle, Saale, Germany
[7] Ctr Struct Mass Spectrometry, Kurt Mothes Str 3, D-06120 Halle, Saale, Germany
[8] Phospholipid Res Ctr, Neuenheimer Feld 515, D-69120 Heidelberg, Germany
来源
关键词
Azide; Diazirine; Membrane lipids; Miscibility; Peptide/lipid interactions; Photo-cross-linking; Mass spectrometry; XL-MS; PHASE-TRANSITIONS; FATTY-ACIDS; BILAYER; PHOSPHOLIPIDS; PROTEINS; CHOLESTEROL; PHOTOLYSIS; MOLECULES; BEHAVIOR; WALP;
D O I
10.1016/j.bbamem.2022.184004
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Although the incorporation of photo-activatable lipids into membranes potentially opens new avenues for studying interactions with peptides and proteins, the question of whether azide- or diazirine-modified lipids are suitable for such studies remains controversial. We have recently shown that diazirine-modified lipids can indeed form cross-links to membrane peptides after UV activation and that these cross-links can be precisely determined in their position by mass spectrometry (MS). However, we also observed an unexpected backfolding of the lipid's diazirine-containing stearoyl chain to the membrane interface challenging the potential application of this modified lipid for future cross-linking (XL)-MS studies of protein/lipid interactions. In this work, we compared an azide- (AzidoPC) and a diazirine-modified (DiazPC) membrane lipid regarding their self-assembly properties, their mixing behavior with saturated bilayer-forming phospholipids, and their reactivity upon UV activation using differential scanning calorimetry (DSC), dynamic light scattering (DLS), small-angle X-ray scattering (SAXS), transmission electron microscopy (TEM), and MS. Mixtures of both modified lipids with DMPC were further used for photo-chemically induced XL experiments with a transmembrane model peptide (KLAW23) to elucidate similarities and differences between the azide and the diazirine moiety. We showed that both photo-reactive lipids can be used to study lipid/peptide and lipid/protein interactions. The AzidoPC proved easier to handle, whereas the DiazPC had fewer degradation products and a higher cross-linking yield. However, the problem of backfolding occurs in both lipids; thus, it seems to be a general phenomenon.
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页数:13
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