The specificity of monoglyceride-protein interactions and mechanism of the protein induced L(beta) to coagel phase transition.
This study aims at gaining insight into the specificity and molecular mechanism of monoglyceride-protein interactions. We used beta-lactoglobulin (beta-LG) and lysozyme as model proteins and both monostearoylglycerol and monopalmitoylglycerol as defined gel phase monoglycerides. The monoglycerides were used in different combinations with the two negatively charged amphiphiles dicetylphosphate and distearylphosphate. The interactions were characterized using the monolayer technique, isothermal titration calorimetry, (2)H-nuclear magnetic resonance (NMR) using deuterium labelled monoglycerides and freeze fracture electron microscopy (EM). Our results show that lysozyme inserts efficiently into all monolayers tested, including pure monoglyceride layers. The insertion of beta-LG depends on the lipid composition of the monolayer and is promoted when the acylchains of the negatively charged amphiphile are shorter than that of the monoglyceride. The binding parameters found for the interaction of beta-LG and lysozyme with monoglyceride bilayers were generally similar. Moreover, in all cases a large exothermic binding enthalpy was observed which was found to depend on the nature of the monoglycerides but not of the proteins. (2)H-NMR and freeze fracture EM showed that this large enthalpy results from a protein mediated catalysis of the monoglyceride L(beta) to coagel phase transition. The mechanism of this phase transition consists of two steps, an initial protein mediated vesicle aggregation step which is followed by stacking and probably fusion of the bilayers.
Boots
J P
JP
Department of Biochemistry of Membranes, Centre for Biomembranes and Lipid Enzymology, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. j.w.p.boots@chem.uu.nl
Chupin
V
V
Killian
J A
JA
Demel
R A
RA
de Kruijff
B
B
D005614:Freeze Fracturing; D005989:Glycerides / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry* / Q000235:genetics; D008051:Lipid Bilayers / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008854:Microscopy, Electron; D008956:Models, Chemical; D008968:Molecular Conformation; D009113:Muramidase / Q000737:chemistry*; D010755:Organophosphates / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D013499:Surface Properties; D013696:Temperature; D013816:Thermodynamics
D005989:Glycerides; D007782:Lactoglobulins; D008051:Lipid Bilayers; D010755:Organophosphates; D011506:Proteins; C002290:dicetylphosphate; D009113:Muramidase