2-DE using hemi-fluorinated surfactants.
The synthesis of hemi-fluorinated zwitterionic surfactants was realized and assessed for 2-DE, a powerful separation method for proteomic analysis. These new fluorinated amidosulfobetaine (FASB-p,m) were compared to their hydrocarbon counterparts amidosulfobetaine (ASB-n) characterized by a hydrophilic polar head, a hydrophobic and lipophilic tail, and an amido group as connector. The tail of these FASB surfactants was in part fluorinated resulting in the modulation of its lipophilicity (or oleophobicity). Their effect on the red blood cell (RBC) membrane showed a specific solubilization depending on the length of the hydrophobic part. A large number of polypeptide spots appeared in the 2-DE patterns by using FASB-p,m. The oleophobic character of these surfactants was confirmed by the fact that Band 3, a highly hydrophobic transmembrane protein, was not solubilized by these fluorinated structures. The corresponding pellet was very rich in Band 3 and could then be solubilized by using a strong detergent such as amidosulfobetaine with an alkyl tail containing 14 carbon atoms (ASB-14). Thus, these hemi-fluorinated surfactants appeared as powerful tools when used at the first step of a two-step solubilization strategy using a hydrocarbon homologous surfactant in the second step.
Starita-Geribaldi
Mireille
M
Connexines et Prolifération Germinale: Physiopathologie Cellulaire et Moléculaire (INSERM U670), UFR de Médecine, Université de Nice Sophia-Antipolis, Nice, France. mgeribal@unice.fr
Thebault
Pascal
P
Taffin de Givenchy
Elisabeth
E
Guittard
Frederic
F
Geribaldi
Serge
S
D001622:Betaine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004910:Erythrocyte Membrane / Q000737:chemistry; D004912:Erythrocytes / Q000737:chemistry; D006801:Humans; D006845:Hydrocarbons, Fluorinated / Q000138:chemical synthesis / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012995:Solubility; D013501:Surface-Active Agents / Q000138:chemical synthesis / Q000737:chemistry*
D006845:Hydrocarbons, Fluorinated; D008565:Membrane Proteins; D013501:Surface-Active Agents; C506587:amidosulfobetaine-14; D001622:Betaine