Salt effects on protein stability: two-stranded alpha-helical coiled-coils containing inter- or intrahelical ion pairs.

An investigation into the role of surface-accessible ion pairs in protein stability was carried out by determining the effects of added salt (KCl, MgCl2 and LaCl3) at neutral and acidic pH on the stability of de novo designed two-stranded alpha-helical coiled-coils. The effects of salt on the stability of coiled-coils containing interhelical i to i' + 5 or intrahelical i to i + 4 and i to i + 3 Lys-Glu ion pairs were compared to the effects on the stability of a control coiled-coil, which contained no intra- or interhelical ion pair. These studies indicate that ionic interactions contribute to coiled-coil stability. The results show that added salt can have complex effects on protein stability, involving stabilizing and destabilizing contributions with the net effect depending on the nature of the charged residues and ionic interactions present in the protein.

[1]  B. Tidor,et al.  Do salt bridges stabilize proteins? A continuum electrostatic analysis , 1994, Protein science : a publication of the Protein Society.

[2]  D A Parry,et al.  Alpha-helical coiled coils: more facts and better predictions. , 1994, Science.

[3]  R. Hodges,et al.  Effect of chain length on the formation and stability of synthetic alpha-helical coiled coils. , 1994, Biochemistry.

[4]  R. Hodges,et al.  Design, synthesis and structural characterization of model heterodimeric coiled-coil proteins. , 2009, International journal of peptide and protein research.

[5]  T. Richmond,et al.  The X-ray structure of the GCN4-bZIP bound to ATF/CREB site DNA shows the complex depends on DNA flexibility. , 1993, Journal of molecular biology.

[6]  A. Mclachlan,et al.  Tropomyosin coiled-coil interactions: evidence for an unstaggered structure. , 1975, Journal of molecular biology.

[7]  I. Chaiken,et al.  Controlled formation of model homo- and heterodimer coiled coil polypeptides. , 1993, Biochemistry.

[8]  R. Hodges,et al.  Protein destabilization by electrostatic repulsions in the two‐stranded α‐helical coiled‐coil/leucine zipper , 1995, Protein science : a publication of the Protein Society.

[9]  R. Hodges,et al.  Electrostatic interactions control the parallel and antiparallel orientation of alpha-helical chains in two-stranded alpha-helical coiled-coils. , 1994, Biochemistry.

[10]  R. L. Baldwin,et al.  How Hofmeister ion interactions affect protein stability. , 1996, Biophysical journal.

[11]  K. Struhl,et al.  The GCN4 basic region leucine zipper binds DNA as a dimer of uninterrupted α Helices: Crystal structure of the protein-DNA complex , 1992, Cell.

[12]  K. Struhl,et al.  Dimerization of leucine zippers analyzed by random selection. , 1993, Nucleic acids research.

[13]  C. Vinson,et al.  Dimerization specificity of the leucine zipper-containing bZIP motif on DNA binding: prediction and rational design. , 1993, Genes & development.

[14]  U. Sauer,et al.  Contributions of engineered surface salt bridges to the stability of T4 lysozyme determined by directed mutagenesis. , 1991, Biochemistry.

[15]  P. S. Kim,et al.  Secondary structure of a leucine zipper determined by nuclear magnetic resonance spectroscopy. , 1990, Biochemistry.

[16]  P. Lyu,et al.  Energetic contribution of solvent-exposed ion pairs to alpha-helix structure. , 1992, Journal of molecular biology.

[17]  A. Lupas,et al.  Predicting coiled coils from protein sequences , 1991, Science.

[18]  A. Soper,et al.  Effect of high salt concentrations on water structure , 1995, Nature.

[19]  P. S. Kim,et al.  Mechanism of specificity in the Fos-Jun oncoprotein heterodimer , 1992, Cell.

[20]  G. S. Manning.  On the application of polyelectrolyte “limiting laws” to the helix‐coil transition of DNA. I. Excess univalent cations , 1972, Biopolymers.

[21]  E. Stellwagen,et al.  The contribution of residue ion pairs to the helical stability of a model peptide , 1992, Biopolymers.

[22]  A. Baxevanis,et al.  Interactions of coiled coils in transcription factors: where is the specificity? , 1993, Current opinion in genetics & development.

[23]  R. Hodges,et al.  Tropomyosin: a model protein for studying coiled-coil and .alpha.-helix stabilization , 1982 .

[24]  T. Arakawa,et al.  Why preferential hydration does not always stabilize the native structure of globular proteins. , 1990, Biochemistry.

[25]  Unzipping the secrets of coiled-coils , 1992, Current Biology.

[26]  R. Hodges,et al.  Structure, function and application of the coiled-coil protein folding motif. , 1993, Current opinion in biotechnology.

[27]  P. S. Kim,et al.  Measurement of interhelical electrostatic interactions in the GCN4 leucine zipper. , 1995, Science.

[28]  Serge N. Timasheff,et al.  Preferential interactions determine protein solubility in three-component solutions: the magnesium chloride system , 1990 .

[29]  R. Hodges,et al.  Disulfide bond contribution to protein stability: positional effects of substitution in the hydrophobic core of the two-stranded alpha-helical coiled-coil. , 1993, Biochemistry.

[30]  R. L. Baldwin,et al.  The energetics of ion-pair and hydrogen-bonding interactions in a helical peptide. , 1993, Biochemistry.

[31]  R. Hodges,et al.  Tropomyosin: Amino Acid Sequence and Coiled-Coil Structure , 1973 .

[32]  R. Hodges,et al.  The role of interhelical ionic interactions in controlling protein folding and stability. De novo designed synthetic two-stranded alpha-helical coiled-coils. , 1994, Journal of molecular biology.

[33]  S. Sligar,et al.  Electrostatic stabilization in four‐helix bundle proteins , 1993, Protein science : a publication of the Protein Society.

[34]  T. Gibson,et al.  The solution structure of a leucine-zipper motif peptide. , 1991, Protein engineering.

[35]  D. Shortle,et al.  Contributions of the polar, uncharged amino acids to the stability of staphylococcal nuclease: evidence for mutational effects on the free energy of the denatured state. , 1992, Biochemistry.

[36]  L. Bracco,et al.  Construction, purification, and characterization of a hybrid protein comprising the DNA binding domain of the LexA repressor and the Jun leucine zipper: a circular dichroism and mutagenesis study. , 1991, Biochemistry.

[37]  S. N. Timasheff,et al.  On the role of surface tension in the stabilization of globular proteins , 1996, Protein science : a publication of the Protein Society.

[38]  R. Hodges,et al.  The net energetic contribution of interhelical electrostatic attractions to coiled-coil stability. , 1994, Protein engineering.

[39]  P. S. Kim,et al.  Evidence that the leucine zipper is a coiled coil. , 1989, Science.

[40]  F. Crick,et al.  The packing of α‐helices: simple coiled‐coils , 1953 .

[41]  G. S. Manning.  The molecular theory of polyelectrolyte solutions with applications to the electrostatic properties of polynucleotides , 1978, Quarterly Reviews of Biophysics.

[42]  A. Fersht,et al.  Estimating the contribution of engineered surface electrostatic interactions to protein stability by using double-mutant cycles. , 1990, Biochemistry.

[43]  P. S. Kim,et al.  Interhelical Salt Bridges, Coiled-Coil Stability, and Specificity of Dimerization , 1996, Science.

[44]  R. Sauer,et al.  The Basic-Region Leucine-Zipper Family of DNA Binding Proteins , 1992 .

[45]  S. Lowey.  COMPARATIVE STUDY OF THE ALPHA-HELICAL MUSCLE PROTEINS. TYROSYL TITRATION AND EFFECT OF PH ON CONFORMATION. , 1965, Journal of Biological Chemistry.

[46]  C. Mant,et al.  Reversed-phase liquid chromatography as a useful probe of hydrophobic interactions involved in protein folding and protein stability. , 1994, Journal of chromatography. A.

[47]  O. Ptitsyn,et al.  Determination of stability of the DNA double helix in an aqueous medium , 1969 .

[48]  R. Hodges,et al.  Protein denaturation with guanidine hydrochloride or urea provides a different estimate of stability depending on the contributions of electrostatic interactions , 1994, Protein science : a publication of the Protein Society.

[49]  R. Hodges,et al.  Synthesis of a model protein of defined secondary and quaternary structure. Effect of chain length on the stabilization and formation of two-stranded alpha-helical coiled-coils. , 1984, The Journal of biological chemistry.

[50]  R. Hodges,et al.  The Effects of Interhelical Electrostatic Repulsions between Glutamic Acid Residues in Controlling the Dimerization and Stability of Two-stranded α-Helical Coiled-coils (*) , 1995, The Journal of Biological Chemistry.

[51]  R. Hodges,et al.  The two‐stranded α‐helical coiled‐coil is an ideal model for studying protein stability and subunit interactions , 1992, Biopolymers.

[52]  P. S. Kim,et al.  Peptide ‘Velcro’: Design of a heterodimeric coiled coil , 1993, Current Biology.

[53]  R. Hodges,et al.  Investigation of electrostatic interactions in two-stranded coiled-coils through residue shuffling. , 1996, Biophysical chemistry.

[54]  C. Pace.  Determination and analysis of urea and guanidine hydrochloride denaturation curves. , 1986, Methods in enzymology.

[55]  R. Hodges,et al.  Synthetic model proteins: contribution of hydrophobic residues and disulfide bonds to protein stability. , 1990, Peptide research.

[56]  C. Vinson,et al.  A thermodynamic scale for leucine zipper stability and dimerization specificity: e and g interhelical interactions. , 1994, The EMBO journal.

[57]  R. Hodges,et al.  Comparison of antiparallel and parallel two-stranded alpha-helical coiled-coils. Design, synthesis, and characterization. , 1993, The Journal of biological chemistry.

[58]  A. Fink,et al.  Mechanism of acid-induced folding of proteins. , 1990, Biochemistry.

[59]  P. S. Kim,et al.  X-ray structure of the GCN4 leucine zipper, a two-stranded, parallel coiled coil. , 1991, Science.

[60]  J. N. Mark Glover,et al.  Crystal structure of the heterodimeric bZIP transcription factor c-Fos–c-Jun bound to DNA , 1995, Nature.

[61]  B. Honig,et al.  Destabilization of an alpha-helix-bundle protein by helix dipoles. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[62]  R. Hodges,et al.  Synthetic model proteins: the relative contribution of leucine residues at the nonequivalent positions of the 3-4 hydrophobic repeat to the stability of the two-stranded alpha-helical coiled-coil. , 1992, Biochemistry.

[63]  R. Hodges,et al.  The relative positions of alanine residues in the hydrophobic core control the formation of two-stranded or four-stranded alpha-helical coiled-coils. , 1996, Protein engineering.

[64]  R. Hodges,et al.  Packing and hydrophobicity effects on protein folding and stability: Effects of β‐branched amino acids, valine and isoleucine, on the formation and stability of two‐stranded α‐helical coiled coils/leucine zippers , 1993, Protein science : a publication of the Protein Society.

[65]  James C. Hu,et al.  Sequence requirements for coiled-coils: analysis with lambda repressor-GCN4 leucine zipper fusions. , 1990, Science.

[66]  R. L. Baldwin,et al.  Helix propensities of the amino acids measured in alanine‐based peptides without helix‐stabilizing side‐chain interactions , 1994, Protein science : a publication of the Protein Society.

[67]  R. Müller,et al.  Non-leucine residues in the leucine repeats of Fos and Jun contribute to the stability and determine the specificity of dimerization. , 1991, Nucleic acids research.

[68]  A calorimetric characterization of the salt dependence of the stability of the GCN4 leucine zipper , 1995, Protein science : a publication of the Protein Society.