HPTLC Data of "Metal Ion Cofactors Modulate Integral Enzyme Activity By Varying Differential Membrane Curvature Stress"

Lipid hydrolysis by the integral membrane protein OmpLA (outer membrane phospholipase). The enzymatic degradation of the proteoliposomes was determined by TLC. After lipid extraction against organic solvent (2:1 vol/vol chloroform/methanol) based on the Folch extraction method, the samples were spotted on a silica plate (Sigma-Aldrich, Steinheim, Germany) with the automatic TLC sampler 4 (CAMAG, Muttenz, Switzerland). The mobile phase in the developing chamber was a solvent mixture composed of 32.5:12.5:2 vol/vol/vol CHCl_3/MeOH/H_2O. After drying, the plate was immersed in a developing bath (5.08 g MnCl_2 dissolved in 480 ml H_2O, 480 ml EtOH and 32 ml H_2SO_4), which is sensitive to double bonds, and dried for 15 min at 120°C. To quantify the lipid concentrations the plate was scanned with the TLC scanner 3 (CAMAG, Muttenz, Switzerland) and further analyzed with WinCats software.

Lipids:

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE)

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG)