Effect of the Surface Charge of Artificial Chaperones on the Refolding of Thermally Denatured Lysozymes.

Artificial chaperones are of great interest in fighting protein misfolding and aggregation for the protection of protein bioactivity. A comprehensive understanding of the interaction between artificial chaperones and proteins is critical for the effective utilization of these materials in biomedicine. In this work, we fabricated three kinds of artificial chaperones with different surface charges based on mixed-shell polymeric micelles (MSPMs), and investigated their protective effect for lysozymes under thermal stress. It was found that MSPMs with different surface charges showed distinct chaperone-like behavior, and the neutral MSPM with PEG shell and PMEO2MA hydrophobic domain at high temperature is superior to the negatively and positively charged one, because of the excessive electrostatic interactions between the protein and charged MSPMs. The results may benefit to optimize this kind of artificial chaperone with enhanced properties and expand their application in the future.

[1]  D. Thirumalai,et al.  Chaperonin-mediated protein folding. , 2001, Annual review of biophysics and biomolecular structure.

[2]  Jie Ren,et al.  Synthesis, characterization, and properties of tunable thermosensitive amphiphilic dendrimer‐star copolymers with Y‐shaped arms , 2011 .

[3]  J. Sambrook,et al.  Protein folding in the cell , 1992, Nature.

[4]  T. Ishizone,et al.  Synthesis of Thermally Sensitive Water-Soluble Polymethacrylates by Living Anionic Polymerizations of Oligo(ethylene glycol) Methyl Ether Methacrylates , 2003 .

[5]  Linqi Shi,et al.  In vivo biodistribution of mixed shell micelles with tunable hydrophilic/hydrophobic surface. , 2013, Biomacromolecules.

[6]  Zhenkun Zhang,et al.  Artificial chaperones based on mixed shell polymeric micelles: insight into the mechanism of the interaction of the chaperone with substrate proteins using Förster resonance energy transfer. , 2015, ACS applied materials & interfaces.

[7]  J. Buchner,et al.  Molecular Chaperones — Cellular Machines for Protein Folding , 2002 .

[8]  K. Akiyoshi,et al.  Thermoresponsive controlled association of protein with a dynamic nanogel of hydrophobized polysaccharide and cyclodextrin: heat shock protein-like activity of artificial molecular chaperone. , 2005, Biomacromolecules.

[9]  K. Wüthrich,et al.  GroEL‐GroES‐mediated protein folding , 2006, Chemical reviews.

[10]  A. Sharma,et al.  Cyclodextrins as protein folding aids. , 1995, Biochemical and biophysical research communications.

[11]  Hideki Taguchi,et al.  Single-molecule observation of protein–protein interactions in the chaperonin system , 2001, Nature Biotechnology.

[12]  C. O. Fágáin,et al.  Understanding and increasing protein stability. , 1995, Biochimica et biophysica acta.

[13]  Helen R Saibil,et al.  The Chaperonin ATPase Cycle: Mechanism of Allosteric Switching and Movements of Substrate-Binding Domains in GroEL , 1996, Cell.

[14]  Keitaro Yoshimoto,et al.  Regulation of lysozyme activity based on thermotolerant protein/smart polymer complex formation. , 2009, Journal of the American Chemical Society.

[15]  V. Rotello,et al.  Synthetic "chaperones": nanoparticle-mediated refolding of thermally denatured proteins. , 2008, Chemical communications.

[16]  C. Dobson,et al.  Protein aggregation and aggregate toxicity: new insights into protein folding, misfolding diseases and biological evolution , 2003, Journal of Molecular Medicine.

[17]  Christian Lange,et al.  l‐Arginine increases the solubility of unfolded species of hen egg white lysozyme , 2005, Protein science : a publication of the Protein Society.

[18]  Linqi Shi,et al.  Maintenance of amyloid β peptide homeostasis by artificial chaperones based on mixed-shell polymeric micelles. , 2014, Angewandte Chemie.

[19]  Edmond Magner,et al.  Proteins in mesoporous silicates. , 2008, Angewandte Chemie.

[20]  Theodore W Randolph,et al.  Surfactant-protein interactions. , 2002, Pharmaceutical biotechnology.

[21]  J. Weissman,et al.  Characterization of the Active Intermediate of a GroEL–GroES-Mediated Protein Folding Reaction , 1996, Cell.

[22]  S. Fujiwara,et al.  Biophysical effect of amino acids on the prevention of protein aggregation. , 2002, Journal of biochemistry.

[23]  H. Saibil,et al.  GroEL/GroES‐mediated protein folding , 2006 .

[24]  T. A. Hatton,et al.  Protein refolding in reversed micelles , 1990, Biotechnology and bioengineering.

[25]  Kevin L. Shaw,et al.  Charge–charge interactions influence the denatured state ensemble and contribute to protein stability , 2000, Protein science : a publication of the Protein Society.

[26]  F. Hartl,et al.  Molecular chaperones in cellular protein folding. , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[27]  Junyang Shen,et al.  Controlled release of ionic drugs from complex micelles with charged channels. , 2012, Biomacromolecules.

[28]  J. Finch,et al.  Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA , 1978, Nature.

[29]  C. Pace,et al.  Protein Ionizable Groups: pK Values and Their Contribution to Protein Stability and Solubility* , 2009, Journal of Biological Chemistry.

[30]  George I Makhatadze,et al.  Protein stability and surface electrostatics: a charged relationship. , 2006, Biochemistry.

[31]  Yang Liu,et al.  Temperature-responsive mixed-shell polymeric micelles for the refolding of thermally denatured proteins. , 2013, Chemistry.

[32]  Arthur L Horwich,et al.  Chaperonin-mediated protein folding: using a central cavity to kinetically assist polypeptide chain folding , 2009, Quarterly Reviews of Biophysics.

[33]  A. Minton,et al.  Protein aggregation in crowded environments , 2006, Biological chemistry.

[34]  F. Hartl,et al.  Converging concepts of protein folding in vitro and in vivo , 2009, Nature Structural &Molecular Biology.

[35]  J. B. Jones,et al.  Chemical modification of enzymes for enhanced functionality. , 1999, Current opinion in biotechnology.

[36]  F. Hartl,et al.  Molecular Chaperones in the Cytosol: from Nascent Chain to Folded Protein , 2002, Science.

[37]  S. Lindquist,et al.  Multiple effects of trehalose on protein folding in vitro and in vivo. , 1998, Molecular cell.

[38]  James R. Swartz,et al.  Polyethylene Glycol Enhanced Protein Refolding , 1992, Bio/Technology.

[39]  G. Vriend,et al.  Engineering an enzyme to resist boiling. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Michele Vendruscolo,et al.  Protein folding and misfolding: a paradigm of self–assembly and regulation in complex biological systems , 2003, Philosophical Transactions of the Royal Society of London. Series A: Mathematical, Physical and Engineering Sciences.

[41]  Ángel Berenguer-Murcia,et al.  Potential of Different Enzyme Immobilization Strategies to Improve Enzyme Performance , 2011 .

[42]  F. Hartl,et al.  Protein folding in the central cavity of the GroEL–GroES chaperonin complex , 1996, Nature.

[43]  Andrew D. Miller,et al.  The mechanism of GroEL/GroES folding/refolding of protein substrates revisited. , 2006, Organic & biomolecular chemistry.

[44]  C. Dobson.  Protein folding and misfolding , 2003, Nature.

[45]  S. Gellman,et al.  Artificial chaperones-protein refolding via sequential use of detergent and cyclodextrin , 1995 .

[46]  P. Pedersen,et al.  Defective protein folding as a basis of human disease. , 1995, Trends in biochemical sciences.

[47]  M. Reetz,et al.  An artificial metalloenzyme: creation of a designed copper binding site in a thermostable protein. , 2010, Angewandte Chemie.

[48]  K. Dill,et al.  Protein stability: electrostatics and compact denatured states. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[49]  A. Bhaw-Luximon,et al.  Amphiphilic Poly(L-lysine-b-caprolactone) Block Copolymers : Synthesis, Characterization, and Solution Properties , 2008 .

[50]  G. Makhatadze,et al.  Engineering a thermostable protein via optimization of charge-charge interactions on the protein surface. , 1999, Biochemistry.

[51]  Lucia Gardossi,et al.  Understanding enzyme immobilisation. , 2009, Chemical Society reviews.

[52]  G. L. Butterfoss,et al.  Structural and functional studies of Aspergillus oryzae cutinase: enhanced thermostability and hydrolytic activity of synthetic ester and polyester degradation. , 2009, Journal of the American Chemical Society.

[53]  P. Horowitz,et al.  Detergent-assisted refolding of guanidinium chloride-denatured rhodanese. The effect of lauryl maltoside. , 1986, The Journal of biological chemistry.

[54]  Colin Norman,et al.  What Don't We Know? , 2005, Science.