title: Measurement and control of pH in the aqueous interior of reverse micelles.
abstract: The encapsulation of proteins and nucleic acids within the nanoscale water core of reverse micelles has been used for over 3 decades as a vehicle for a wide range of investigations including enzymology, the physical chemistry of confined spaces, protein and nucleic acid structural biology, and drug development and delivery. Unfortunately, the static and dynamical aspects of the distribution of water in solutions of reverse micelles complicate the measurement and interpretation of fundamental parameters such as pH. This is a severe disadvantage in the context of (bio)chemical reactions and protein structure and function, which are generally highly sensitive to pH. There is a need to more fully characterize and control the effective pH of the reverse micelle water core. The buffering effect of titratable head groups of the reverse micelle surfactants is found to often be the dominant variable defining the pH of the water core. Methods for measuring the pH of the reverse micelle aqueous interior using one-dimensional (1)H and two-dimensional heteronuclear NMR spectroscopy are described. Strategies for setting the effective pH of the reverse micelle water core are demonstrated. The exquisite sensitivity of encapsulated proteins to the surfactant, water content, and pH of the reverse micelle is also addressed. These results highlight the importance of assessing the structural fidelity of the encapsulated protein using multidimensional NMR before embarking upon a detailed structural and biophysical characterization.
authors: Graduate Group in Biochemistry and Molecular Biophysics and Johnson Research Foundation and Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine , Philadelphia, Pennsylvania 19104-6059, United States.
authors: Bryan S
authors: BS
authors: Marques
authors: Nathaniel V
authors: NV
authors: Nucci
authors: Igor
authors: I
authors: Dodevski
authors: Kristina W C
authors: KW
authors: Wang
authors: Evangelia A
authors: EA
authors: Athanasoula
authors: Christine
authors: C
authors: Jorge
authors: A Joshua
authors: AJ
authors: Wand
mesh_terms: D006801:Humans; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry; D013997:Time Factors; D014867:Water / Q000737:chemistry*
chemical_list: D008823:Micelles; D011506:Proteins; D013501:Surface-Active Agents; D014867:Water