chloroform dissolved e coli polar lipid extract  (Croda International Plc)

Journal: Channels (Austin, Tex.)

Article Title: Effects of lipid environment on the conformational changes of an ABC importer.

doi: 10.4161/chan.29294

Figure Lengend Snippet: Figure 1. Chemical properties of detergents and lipids. (A) n-Decyl-β-D- maltopyranoside is a common non-ionic detergent used to stabilize TM proteins.5 The large polar head group and relatively short non-polar tail give the detergent a roughly conical shape, which facilitates the formation of detergent micelles. (B) Phosphatidylethanolamine can represent up to 70% of lipid in bacterial membranes;35,46 its roughly cylindrical shape enables the formation of lipid bilay- ers. (C) In addition to triglycerides and hopanoids, phospholipids are an important component of lipid bilayers. Phospholipids contain three components attached to a glycerol backbone: two non-polar tails (R, light gray) and one polar head group (X, dark gray). Here we show a selection of head groups including cardiolipin (CA), phosphatidylglycerol (PG), phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI); and major tail groups from E. coli: palmitic acid (C16:0), oleic acid (C18:1), linoleic acid (C18:2), 9,10-methylene-hexadecanoic acid (cyclopropyl C17:0), and lactobacillic acid (cyclopropyl C19:0). Chemical structures were prepared using ChemDraw (CambridgeSoft Inc.).

Article Snippet: Lipid preparation Chloroform-dissolved E. coli Polar lipid (Avanti: 100600C) and Egg PC (Avanti: 840051C) were mixed in a 3:1 (w:w) ratio.

Techniques: Selection

Journal: Channels (Austin, Tex.)

Article Title: Effects of lipid environment on the conformational changes of an ABC importer.

doi: 10.4161/chan.29294

Figure Lengend Snippet: Figure 2. CW-EPR spectroscopy of MolBC_N93C+MTSL and I151C+MTSL showing the effect of lipid reconstitution on TMD conformation change. (A) Ribbon and space fill diagram of MolBC with N93 and I151 shown in black spheres. CW-EPR spectra of MolBC_N93C+MTSL (B-D) and I151C+MTSL (E-G) stabilized in E. coli polar lipid: Egg PC (3:1) liposomes or DM detergent micelles (black or gray respectively). Room temperature spectra (250 Gauss scan width) were recorded apo (B and E), ATP-bound (C and F), and post-hydrolysis (D and G). N93C spectra were normalized by the height of the central peak. I151C spectra were normalized for equal spin (normalized double integration values). MolBC surface and ribbon diagrams were prepared using PyMOL.47 Spectra were graphed using Grapher 9 (Golden Software Inc.).

Article Snippet: Lipid preparation Chloroform-dissolved E. coli Polar lipid (Avanti: 100600C) and Egg PC (Avanti: 840051C) were mixed in a 3:1 (w:w) ratio.

Techniques: Spectroscopy, Liposomes, Software

Journal: Channels (Austin, Tex.)

Article Title: Effects of lipid environment on the conformational changes of an ABC importer.

doi: 10.4161/chan.29294

Figure Lengend Snippet: Figure 3. Effect of lipid identity on nucleotide-dependent conformation change, shown by CW-EPR. (A) MolB_N93C and MolBC_I151C were spin-labeled with MTSL then recon- stituted into soybean PC liposomes (red), E. coli polar lipid: Egg PC (3:1) liposomes (black), or E. coli total lipid liposomes (blue). CW-EPR spectra (250G) were recorded apo, ATP- bound, and post-hydrolysis. For N93C, spectra from samples in E. coli polar: Egg PC are overlaid with soybean PC (A) or E. coli total lipid spectra (B) and were normalized by the height of the central peak. (C) For I151C, the E. coli polar: Egg PC spectra are overlaid with soybean PC and were normalized for equal spin (normalized double integration values).

Article Snippet: Lipid preparation Chloroform-dissolved E. coli Polar lipid (Avanti: 100600C) and Egg PC (Avanti: 840051C) were mixed in a 3:1 (w:w) ratio.

Techniques: Labeling, Liposomes