Pulmonary surfactant protein SP‐B is significantly more immunoreactive in anionic than in zwitterionic bilayers

[摘要]

Binding of polyclonal and monoclonal antibodies, quantitated by enzyme-linked immunosorbent assay, to porcine SP-B reconstituted in different phospholipid bilayers has been used to assess differences in protein structure due to lipid–protein interactions. SP-B bound significantly more antibodies when it was reconstituted in bilayers made of anionic phospholipids (phosphatidic acid, cardiolipin, phosphatidylglycerol, phosphatidylinositol or phosphatidylserine) than in zwitterionic bilayers (phosphatidylcholine, phosphatidylcholine/cholesterol, or phosphatidylethanolamine) or in fatty acid micelles (made of salts of palmitic or stearic acids). These differences in immunoreactivity can be important in the development of quantitation methods for SP-B in clinical samples based on immunological techniques.

[学科分类] 生物化学/生物物理

[关键词] Protein epitope;Enzyme-linked immunosorbent assay;Membrane protein conformation;Surface activity;Lipid–protein interaction;Ch;cholesterol;Chl;chloroform;DMAB;dimethyl aminobenzoic acid;DPG;bovine heart diphosphatidylglycerol;DPPC;1;2-dipalmitoyl-sn-glycero-3-phosphocholine;DPPG;1;2-dipalmitoyl-sn-glycero-3-phospho-rac-glycerol;ECL;enzyme chemiluminescence;ELISA;enzyme-linked immunosorbent assay;LPC;1-palmitoyl-2-lyso-sn-glycero-3-phosphocholine;MBTH;3-methyl-2-benzothiazolinone hydrazine;MeOH;methanol;PAGE;polyacrylamide gel electrophoresis;PA;egg yolk phosphatidic acid;Pal;palmitic acid;PC;egg yolk phosphatidylcholine;PE;egg yolk phosphatidylethanolamine;PI;bovine liver phosphatidylinositol;PG;egg yolk phosphatidylglycerol;PS;bovine brain phosphatidylserine;SDS;sodium dodecyl sulfate;SP-A;surfactant protein A;SP-B;surfactant protein B;SP-C;surfactant protein C;Ste;stearic acid