Localizing the Membrane Binding Region of Group VIA Ca2+-independent Phospholipase A2 Using Peptide Amide Hydrogen/Deuterium Exchange Mass Spectrometry

Nuclear Receptors and Their Selective Pharmacologic Modulators

“…This technique has provided complementary information to that gained by X-ray crystallography (Hamuro et al, 2006;Chandra et al, 2008;Hsu et al, 2009;Iacob et al, 2009). HDX was used to probe the conformational dynamics of the LBD of VDR in complex with three ligands, one being 1,25-(OH) 2 D 3 .…”

Structural basis of specific interactions of Lp-PLA2 with HDL revealed by hydrogen deuterium exchange mass spectrometry

“…D 2 O buffer consisted of 12 mM Tris, 50 mM NaCl in 99% D 2 O, pD read 7.5, as previously described (31)(32)(33)(34)(35). Exchange experiments were initiated by mixing 20 µ L of Lp-PLA 2 (2.5 mg/ml) in protein buffer, or Lp-PLA 2 preincubated with human HDL, apoA-I, apoA-II, or BSA with 60 µ L of D 2 O buffer to a fi nal concentration of 50% D 2 O.…”

Roles of Acidic Phospholipids and Nucleotides in Regulating Membrane Binding and Activity of a Calcium-independent Phospholipase A2 Isoform

“…A requirement for coincidence detection of both a protein and acidic phospholipids may provide sufficient stringency to prevent constitutive lipase activity while still enabling rapid changes in GVIA-iPLA 2 localization to control the architecture of multiple organelles. Notably, deuterium exchange experiments were recently conducted using human GVIA-2 to monitor its association with neutral phospholipid membranes (45). It will be interesting to determine how the presence of acidic phospholipids influences GVIA-2 membrane binding and to dissect the regions of GVIA-2 that respond specifically to charged and uncharged lipid headgroups.…”