Interactions of gemini surfactants with two model proteins: NMR, CD, and fluorescence spectroscopies.
Gemini surfactants have two polar head groups and two hydrocarbon tails. Compared with conventional surfactants, geminis have much lower (μM vs. mM) critical micelle concentrations and possess slower (ms vs. μs) monomer <-- / --> micelle kinetics. The structure of the gemini surfactants studied is [HOCH(2)CH(2)-, CH(3)-, CH(3)(CH(2))(15)-N(+)-(CH(2))(s)-N(+)-(CH(2))(15)CH(3),-CH(3),-CH(2)CH(2)OH]·2Br(-) where s=4, 5, or 6. Our objective is to reveal the effect of these cationic gemini surfactants on the structure and stability of two model proteins: Ribonuclease A (RNase A) and Hen Egg White Lysozyme (HEWL). 2D (1)H NMR and Circular Dichroism (CD) spectroscopies show that the conformation of RNase A and HEWL is unaffected at low to neutral pH where these proteins are positively charged, although hydrogen exchange shows that RNase A's conformational stability is slightly lowered. At alkaline pH, where these proteins lose their net positive charge, fluorescence and CD spectroscopies and ITC experiments show that they do interact with gemini surfactants, and multiple protein•gemini complexes are observed. Based on the results, we conclude that these cationic gemini surfactants neither interact strongly with nor severely destabilize these well folded proteins in physiological conditions, and we advance that they can serve as useful membrane mimetics for studying the interactions between membrane components and positively charged proteins.
Department of Chemistry, University of Isfahan, Isfahan, Iran.
Razieh
R
Amiri
Abdol-Khalegh
AK
Bordbar
Ma Flor
MF
García-Mayoral
Ahmad Reza
AR
Khosropour
Iraj
I
Mohammadpoor-Baltork
Margarita
M
Menéndez
Douglas V
DV
Laurents
D000818:Animals; D002645:Chickens; D002942:Circular Dichroism; D009682:Magnetic Resonance Spectroscopy; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013501:Surface-Active Agents / Q000737:chemistry / Q000378:metabolism*; D013816:Thermodynamics
D013501:Surface-Active Agents; D012259:Ribonuclease, Pancreatic; C421864:hen egg lysozyme; D009113:Muramidase