Protein loop compaction and the origin of the effect of arginine and glutamic acid mixtures on solubility, stability and transient oligomerization of proteins

Addition of a 50 mM mixture of l-arginine and l-glutamic acid (RE) is extensively used to improve protein solubility and stability, although the origin of the effect is not well understood. We present Small Angle X-ray Scattering (SAXS) and Nuclear Magnetic Resonance (NMR) results showing that RE induces protein compaction by collapsing flexible loops on the protein core. This is suggested to be a general mechanism preventing aggregation and improving resistance to proteases and to originate from the polyelectrolyte nature of RE. Molecular polyelectrolyte mixtures are expected to display long range correlation effects according to dressed interaction site theory. We hypothesize that perturbation of the RE solution by dissolved proteins is proportional to the volume occupied by the protein. As a consequence, loop collapse, minimizing the effective protein volume, is favored in the presence of RE

Complexes biomimétiques de la superoxyde dismutase

STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF

Effect of Water/Carboxymethylcellulose Gel on the Excimer Formation of Polyamine Ligands Functionalized with Naphthalene

Formation of intramolecular excimers was studied for the compounds 6,20-bis-naphthalene-2-ylmethyl- 3,6,9,17,20,23,29,30-octaaza-tricyclo[23.3.1.1] triaconta-1(29),11,13,15(30),25,27-hexane (L1), a bisnaphthalene derivative, and N1-(2-{bis-[2-(3-amino-propylamino)-ethyl]-amino}-ethyl)-propane-1,3-diamine (L2), a tris-naphthalene derivative, incorporated in gels of carboxymethylcellulose sodium salt. Excimers are formed through dynamic processes as well as from ground state dimers. A mathematical treatment including preformed dimers was used to split the static and dynamic contributions in the excimer/monomer emission ratio. In the case of compound L1, the activation energy for excimer formation in water is 11 kJ mol-1 and experimental evidence that the dynamic terms are identical in water and in the gel was achieved. On the other hand, ground state dimers are extremely favorable in the gel with an equilibrium constant of 8.2 at 25 °C. On the contrary, in the case of compound L2, the ground state dimers are observed in water but not in the gel. The results were interpreted as reflecting a balance between specific interactions (hydrogen bond) and confinement effects

Effects of preorganization in the chelation of UO22+ by hydroxamate ligands: cyclic PIPO– vs linear NMA–

International audienceMany siderophores incorporate as bidentate chelating subunits linear and more seldomly cyclic hydroxamate groups. In this work, a comparative study of the uranyl binding properties in aqueous solution of two monohydroxamic acids, the prototypical linear N-methylacetohydroxamic acid (NMAH) and the cyclic analog 1-hydroxypiperidine-2-one (PIPOH), has been carried out. The complex [UO2(PIPO)(2)(H2O)] crystallized from slightly acidic water solutions (pH < 5), and its molecular structure was determined by X-ray diffraction. The uranyl speciation in the presence of both ligands has been thoroughly investigated in a 0.1 M KNO3 medium at 298.2 K by the combined use of four complementary techniques, i.e., potentiometry, spectrophotometry, Raman spectroscopy, and affinity capillary electrophoresis. Preorganization of the hydroxamate ligand for chelation by incorporation into a cyclic structure, as in PIPO-, results in an increase of nearly one order of magnitude in the formation constants of the uranyl complexes of 1 : 1 and 1 : 2 metal/ligand stoichiometries

Protein loop compaction and the origin of the effect of arginine and glutamic acid mixtures on solubility, stability and transient oligomerization of proteins

Addition of a 50 mM mixture of l-arginine and l-glutamic acid (RE) is extensively used to improve protein solubility and stability, although the origin of the effect is not well understood. We present Small Angle X-ray Scattering (SAXS) and Nuclear Magnetic Resonance (NMR) results showing that RE induces protein compaction by collapsing flexible loops on the protein core. This is suggested to be a general mechanism preventing aggregation and improving resistance to proteases and to originate from the polyelectrolyte nature of RE. Molecular polyelectrolyte mixtures are expected to display long range correlation effects according to dressed interaction site theory. We hypothesize that perturbation of the RE solution by dissolved proteins is proportional to the volume occupied by the protein. As a consequence, loop collapse, minimizing the effective protein volume, is favored in the presence of RE