ABSTRACT
Luminal acid causes intracellular acidification in the gastric epithelium, but the mechanism by which H(+) enters surface cells remains obscure. This study addressed the problem by assessing how different acids affect intracellular pH in gastric surface cells. Isolated Necturus maculosus antral mucosa was exposed to HCl, HNO(3), H(2)SO(4), and H(3)PO(4) at pH 2.30. Intracellular pH was measured with microelectrodes. The physicochemical interaction of a synthetic model of gastric phospholipids with the different acids was studied using Langmuir film balance. Exposure to luminal HNO(3), H(2)SO(4), or H(3)PO(4) caused significantly larger intracellular acidification than exposure to HCl. The degree of acidification was not dependent on the valence or nature of the anionic counterion of the acid but significantly correlated with the amount of molecular acid. By Langmuir film balance, subphases acidified with HNO(3), H(2)SO(4), or H(3)PO(4) caused more close packing of phospholipid molecules than those acidified with HCl, possibly allowing hydrogen bonding between head groups to facilitate H(+) movement across the phospholipid membrane. HCl causes significantly less intracellular acidification in gastric epithelium than HNO(3), H(2)SO(4), or H(3)PO(4). This may be caused by the lower amount of molecular HCl in solution and possible hydrogen bonding between the head groups of phospholipid molecules and the other acids.
Subject(s)
Acids/metabolism , Epithelial Cells/metabolism , Gastric Mucosa/metabolism , Intracellular Fluid/metabolism , Acids/chemistry , Acids/pharmacology , Animals , Diffusion , Electric Impedance , Epithelial Cells/drug effects , Gastric Mucosa/drug effects , Hydrochloric Acid/metabolism , Hydrochloric Acid/pharmacology , Hydrogen-Ion Concentration/drug effects , In Vitro Techniques , Intracellular Fluid/drug effects , Membrane Potentials/drug effects , Necturus , Nitric Acid/chemistry , Nitric Acid/metabolism , Nitric Acid/pharmacology , Phospholipids/chemistry , Phosphoric Acids/chemistry , Phosphoric Acids/metabolism , Phosphoric Acids/pharmacology , Pressure , Sulfuric Acids/chemistry , Sulfuric Acids/metabolism , Sulfuric Acids/pharmacology , Surface Properties/drug effectsABSTRACT
The covalent attachment of Fab' fragments of polyclonal anti-human IgG to a lipid with a terminal linker group was examined by means of quartz crystal microbalance and surface plasmon resonance measurements. The linker lipid was embedded in binary or ternary monolayers of dipalmitoylphosphatidylcholine (DPPC) and cholesterol. Atomic force microscopy images of the films deposited on silanised SiO(2) substrates showed that Fab' fragments take a standing position, thus giving site-directed immobilisation. Human IgG forms a network on interaction with the antibodies. Non-specific binding of bovine serum albumin was found to be very low when DPPC was used as the host matrix. At an optimal Fab' fragment concentration a binding capacity above 60% was obtained. However, if the surface concentration of the immobilised antibodies was too high, the binding capacity decreased due to steric hindrance. The results demonstrate that the covalent coupling of Fab' fragments to N-(epsilon-maleimidocaproyl)-dipalmitoylphosphatidylethanolamine (DPPE-EMC) embedded in a host monolayer matrix of DPPC is a promising approach to achieve a site-directed immobilisation of antibodies with high antigen-binding efficiency.
Subject(s)
Immunoglobulin Fragments/immunology , Lipids/immunology , Surface Plasmon Resonance/methods , 1,2-Dipalmitoylphosphatidylcholine/chemistry , Humans , Immunoglobulin Fab Fragments/immunology , Immunoglobulin Fragments/chemistry , Lipids/chemistry , Microscopy, Atomic Force , Molecular Structure , Phosphatidylethanolamines/chemistryABSTRACT
The layer formation of unilamellar vesicles of L-alpha-dimyristoyl phosphatidylcholine (DMPC) spread onto the air/liquid interface has been investigated. The layers were transferred to clean glass slides and onto slides made hydrophobic with multilayers of Cd arachidate. Aged vesicle suspensions aggregate during storage and are transferred as large domains as imaged with atomic force microscopy (AFM). Freshly prepared vesicles fuse and can be transferred as monolayers to hydrophobic supports. Furthermore, AFM images reveal the importance of positioning the solid support parallel to the moving barrier in order to obtain more uniform deposition of Cd arachidate.