Polymerization mechanism of polypeptide chain aggregation.

The misfolding of polypeptide chains and aggregation into the insoluble inclusion body state is a serious problem for biotechnology and biomedical research. Developing a rational strategy to control aggregation requires understanding the mechanism of polymerization. We investigated the in vitro aggregation of P22 tailspike polypeptide chains by classical light scattering, nondenaturing gel electrophoresis, two-dimensional polyacrylamide gel electrophoresis (PAGE), and computer simulations. The aggregation of polypeptide chains during refolding occurred by multimeric polymerization, in which two multimers of any size could associate to form a larger aggregate and did not require a sequential addition of monomeric subunits. The cluster-cluster polymerization mechanism of aggregation is an important determinant in the kinetic competition between productive folding and inclusion body formation. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 333-343, 1997.

[1]  J. King,et al.  Mutational analysis of protein folding pathways: the P22 tailspike endorhamnosidase. , 1986, Methods in enzymology.

[2]  D A Agard,et al.  Kinetics versus thermodynamics in protein folding. , 1994, Biochemistry.

[3]  G. A. Bowden,et al.  The Effect of Sugars on β‐Lactamase Aggregation in Escherichia coli , 1988 .

[4]  A. Minton,et al.  Confinement as a determinant of macromolecular structure and reactivity. , 1992, Biophysical journal.

[5]  J. King,et al.  Reconstitution of the thermostable trimeric phage P22 tailspike protein from denatured chains in vitro. , 1989, The Journal of biological chemistry.

[6]  R. Jaenicke,et al.  Reconstitution of lactic dehydrogenase. Noncovalent aggregation vs. reactivation. 1. Physical properties and kinetics of aggregation. , 1979, Biochemistry.

[7]  J. King,et al.  Protein Folding Intermediates and Inclusion Body Formation. , 1989, Bio/Technology.

[8]  D L Caspar,et al.  Movement and self-control in protein assemblies. Quasi-equivalence revisited. , 1980, Biophysical journal.

[9]  J. King,et al.  Genetic analysis of the folding pathway for the tail spike protein of phage P22. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. Asakura,et al.  RECONSTITUTION OF BACTERIAL FLAGELLA IN VITRO. , 1964, Journal of molecular biology.

[11]  Daniel I. C. Wang,et al.  Molecular thermodynamic model for Helix‐Helix docking and protein aggregation , 1995 .

[12]  Charles R.scriver,et al.  The Metabolic basis of inherited disease , 1989 .

[13]  C. Frank,et al.  Dynamic light-scattering studies of the fractal aggregation of poly(methacrylic acid) and poly(ethylene glycol) , 1990 .

[14]  B Fane,et al.  Global suppression of protein folding defects and inclusion body formation. , 1991, Science.

[15]  R. Wetzel Mutations and off-pathway aggregation of proteins. , 1994, Trends in biotechnology.

[16]  R. Seckler,et al.  Mechanism of phage P22 tailspike protein folding mutations , 1993, Protein science : a publication of the Protein Society.

[17]  J. King,et al.  Nucleation and growth phases in the polymerization of coat and scaffolding subunits into icosahedral procapsid shells. , 1993, Biophysical journal.

[18]  P. S. Kim,et al.  Intermediates in the folding reactions of small proteins. , 1990, Annual review of biochemistry.

[19]  G. Dollinger,et al.  Practical on-line determination of biopolymer molecular weights by high-performance liquid chromatography with classical light-scattering detection , 1992 .

[20]  R. Seckler,et al.  In vitro folding pathway of phage P22 tailspike protein. , 1991, Biochemistry.

[21]  E. Korn,et al.  The kinetics of actin nucleation and polymerization. , 1983, The Journal of biological chemistry.

[22]  P. Lansbury,et al.  Amyloid fibril formation requires a chemically discriminating nucleation event: studies of an amyloidogenic sequence from the bacterial protein OsmB. , 1992, Biochemistry.

[23]  J. King,et al.  Temperature-sensitive mutants blocked in the folding or subunit assembly of the bacteriophage P22 tail spike protein: II. Active mutant proteins matured at 30 °C , 1981 .

[24]  Daniel I. C. Wang,et al.  Equilibrium Association of a Molten Globule Intermediate in the Refolding of Bovine Carbonic Anhydrase , 1991 .

[25]  J. King,et al.  Formation of aggregates from a thermolabile in vivo folding intermediate in P22 tailspike maturation. A model for inclusion body formation. , 1988, The Journal of biological chemistry.

[26]  J. King,et al.  Thermolabile folding intermediates: inclusion body precursors and chaperonin substrates , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  J. King,et al.  Trimeric intermediate in the in vivo folding and subunit assembly of the tail spike endorhamnosidase of bacteriophage P22. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[28]  J. Hofrichter,et al.  Kinetic studies on photolysis-induced gelation of sickle cell hemoglobin suggest a new mechanism. , 1980, Biophysical journal.

[29]  W. F. Harrington,et al.  Collagen structure in solution. II. Analysis of refolding kinetics in terms of nucleation and growth processes. , 1970, Biochemistry.

[30]  Ken A. Dill,et al.  Aggregation of globular proteins , 1993 .

[31]  S. Katz,et al.  Some problems in particle technology: A statistical mechanical formulation , 1964 .

[32]  Todd M. Przybycien,et al.  Secondary structure characterization of beta-lactamase inclusion bodies. , 1994, Protein engineering.

[33]  G. Georgiou,et al.  Inclusion Bodies and Recovery of Proteins from the Aggregated State , 1991 .

[34]  J. King,et al.  Multimeric intermediates in the pathway to the aggregated inclusion body state for P22 tailspike polypeptide chains , 1995, Protein science : a publication of the Protein Society.

[35]  Robert M. Ziff AGGREGATION KINETICS VIA SMOLUCHOWSKI'S EQUATION , 1984 .

[36]  P. Lansbury,et al.  The carboxy terminus of the beta amyloid protein is critical for the seeding of amyloid formation: implications for the pathogenesis of Alzheimer's disease. , 1993, Biochemistry.

[37]  J. King,et al.  Intracellular trapping of a cytoplasmic folding intermediate of the phage P22 tailspike using iodoacetamide. , 1994, The Journal of biological chemistry.

[38]  J. King,et al.  Temperature-sensitive mutations and second-site suppressor substitutions affect folding of the P22 tailspike protein in vitro. , 1993, The Journal of biological chemistry.

[39]  A. Fink,et al.  Nativelike secondary structure in interleukin-1 beta inclusion bodies by attenuated total reflectance FTIR. , 1994, Biochemistry.

[40]  S. Steinbacher,et al.  Crystal structure of P22 tailspike protein: interdigitated subunits in a thermostable trimer. , 1994, Science.

[41]  B. Vincent,et al.  Coagulation kinetics and structure formation , 1987 .

[42]  D. Brems Solubility of different folding conformers of bovine growth hormone , 1988 .